Instructions For Use: Rotors and Tubes
Instructions For Use: Rotors and Tubes
Instructions For Use: Rotors and Tubes
PN JR-IM-10AG
October 2014
PN JR-IM-10AG iii
Revision History
iv PN JR-IM-10AG
Safety Notice
Read all product manuals and consult with Beckman Coulter-trained personnel before attempting
to operate instrument. Do not attempt to perform any procedure before carefully reading all
instructions. Always follow product labeling and manufacturer’s recommendations. If in doubt as
to how to proceed in any situation, contact your Beckman Coulter Representative.
WARNING
WARNING indicates a potentially hazardous situation which, if not avoided,
could result in death or serious injury.
CAUTION
CAUTION indicates a potentially hazardous situation, which, if not avoided,
may result in minor or moderate injury.
IMPORTANT IMPORTANT is used for comments that add value to the step or procedure being performed.
Following the advice in the Important adds benefit to the performance of a piece of equipment or to a
process.
NOTE NOTE is used to call attention to notable information that should be followed during installation, use,
or servicing of this equipment.
This safety notice summarizes information basic to the safe operation of the rotors and accessories
described in this manual. The international symbol displayed above is a reminder that all safety
instructions should be read and understood before use or maintenance of rotors or accessories.
When you see the symbol on other pages, pay special attention to the safety information presented.
Also observe any safety information contained in applicable rotor and centrifuge manuals.
Observance of safety precautions will help to avoid actions that could cause personal injury, as
well as damage or adversely affect the performance of the centrifuge/rotor/tube system.
Normal operation may involve the use of solutions and test samples that are pathogenic, toxic, or
radioactive. Such materials should not be used in these rotors, however, unless all necessary safety
precautions are taken.
• Observe all cautionary information printed on the original solution containers prior to
their use.
• Handle body fluids with care because they can transmit disease. No known test offers complete
assurance that they are free of micro-organisms. Some of the most virulent—Hepatitis (B and
C) and HIV (I–V) viruses, atypical mycobacteria, and certain systemic fungi—further emphasize
PN JR-IM-10AG v
Safety Notice
Mechanical Safety
the need for aerosol protection. Handle other infectious samples according to good laboratory
procedures and methods to prevent spread of disease. Because spills may generate aerosols,
observe proper safety precautions for aerosol containment. Do not run toxic, pathogenic, or
radioactive materials in the rotor without taking appropriate safety precautions. Biosafe
containment should be used when Risk Group II materials (as identified in the World Health
Organization Laboratory Biosafety Manual) are handled; materials of a higher group require more
than one level of protection.
• Dispose of all waste solutions according to appropriate environmental health and safety
guidelines.
• If disassembly reveals evidence of leakage, you should assume that some fluid escaped the
container or rotor. Apply appropriate decontamination procedures to the centrifuge, rotor,
and accessories.
Mechanical Safety
• Use only the rotors, components, and accessories designed for use in the rotor and centrifuge
being used (refer to the applicable rotor manual). The safety of rotor components and accessories
made by other manufacturers cannot be ascertained by Beckman Coulter. Use of other
manufacturers’ components or accessories in Beckman Coulter rotors may void the rotor warranty
and should be prohibited by your laboratory safety officer.
• Rotors are designed for use at the speeds indicated; however, speed reductions may be required
because of weight considerations of tubes, adapters, and/or the density of the solution being
centrifuged. Be sure to observe the instructions in the applicable rotor manual.
• NEVER attempt to slow or stop a rotor by hand.
• The strength of containers can vary between lots, and will depend on handling and usage. We
highly recommend that you pretest them in the rotor (using buffer or gradient of equivalent
density to the intended sample solution) to determine optimal operating conditions. Scratches
(even microscopic ones) significantly weaken glass and polycarbonate containers.
To help prevent premature failures or hazards by detecting stress corrosion, metal fatigue, wear or
damage to anodized coatings, and to instruct laboratory personnel in the proper care of rotors,
Beckman Coulter offers the Field Rotor Inspection Program (FRIP). This program involves a visit to
your laboratory by a specially trained Beckman Coulter representative, who will inspect all of your
rotors for corrosion or damage. The representative will recommend repair or replacement of at-
risk rotors to prevent potential rotor failures. Contact your local Beckman Coulter office to request
this service.
It is your responsibility to decontaminate the rotors and accessories before requesting service by
Beckman Coulter Field Service.
vi PN JR-IM-10AG
Safety Notice
RoHS Notice
RoHS Notice
These labels and materials declaration table (the Table of Hazardous Substance’s Name and
Concentration) are to meet People’s Republic of China Electronic Industry Standard SJ/T11364-2006
“Marking for Control of Pollution Caused by Electronic Information Products” requirements.
China RoHS Caution Label — This label indicates that the electronic information product contains
certain toxic or hazardous substances. The center number is the Environmentally Friendly Use
Period (EFUP) date, and indicates the number of calendar years the product can be in operation.
Upon the expiration of the EFUP, the product must be immediately recycled. The circling arrows
indicate the product is recyclable. The date code on the label or product indicates the date of
manufacture.
China RoHS Environmental Label — This label indicates that the electronic information product does
not contain any toxic or hazardous substances. The center “e” indicates the product is
environmentally safe and does not have an Environmentally Friendly Use Period (EFUP) date.
Therefore, it can safely be used indefinitely. The circling arrows indicate the product is recyclable.
The date code on the label or product indicates the date of manufacture.
PN JR-IM-10AG vii
Safety Notice
RoHS Notice
viii PN JR-IM-10AG
Contents
Scope, xvii
Scope of this Manual, xvii
Rotors, 1-1
Introduction, 1-1
General Description, 1-1
Rotor Designations, 1-1
Materials, 1-3
Drive Pins, 1-4
Rotor Selection, 1-4
Pelleting (Differential Separation), 1-11
Isopycnic Separations, 1-15
Rate Zonal Separations, 1-15
Blood Component Separations, 1-16
General Operating Information, 1-16
Rotor Balance, 1-16
Rotor Tie-Down, 1-17
Overspeed Protection, 1-18
Allowable Run Speeds, 1-18
Temperature Compensation, 1-19
Avanti J Centrifuges, 1-19
Avanti J2 Series Centrifuges (No longer manufactured), 1-20
Analog J6 Series Centrifuges (No longer manufactured), 1-20
Microprocessor-Controlled Centrifuges, 1-21
ix
Contents
x
Contents
Polypropylene, 3-6
Polyethylene, 3-6
Stainless Steel, 3-6
Capping Tubes, 3-6
Filling and Capping Tubes, 3-7
Three-Piece Assemblies, 3-7
JLA-8.1000 and JLA-9.1000 Bottle Cap/Closure, 3-7
Filling and Loading Cups in the JS-5.0 Rotor, 3-8
Filling and Sealing Quick-Seal Tubes, 3-9
Capping Multiwell Titer Plates, 3-14
Cap Strips, 3-14
Aluminum Foil Lids, 3-14
Using Adapters, 3-14
Using Solid Multitube Adapters, 3-14
Using Modular Disk Multitube Adapters, 3-15
Using Aerosolve Canisters, 3-16
Using Canisters as Wide-Mouth Bottles, 3-17
Using Canisters with Tube Racks, 3-18
Using Blood Bag Cups, 3-19
Sample Recovery, 3-20
Capped Tubes, 3-21
JS-5.0 Cups, 3-21
Quick-Seal Tubes, 3-22
Making Ultra-Clear Tubes Wettable, 3-23
xi
Contents
Using Vertical-Tube
and Rack-Type Rotors, 6-1
Introduction, 6-1
Description, 6-1
Vertical-Tube Rotors, 6-2
Rack-Type Rotors, 6-2
Using a Vertical-Tube Rotor, 6-3
Tubes and Bottles, 6-3
Rotor Preparation and Loading, 6-3
Prerun Safety Checks, 6-3
Rotor Preparation, 6-3
Operation, 6-4
Installing the Rotor, 6-5
Removal and Sample Recovery, 6-6
Using a Rack-Type Rotor, 6-7
Trays and Tubes, 6-7
Rotor Preparation and Loading, 6-7
Prerun Safety Checks, 6-7
Rotor Preparation, 6-7
Operation, 6-8
Installing the Rotor, 6-9
Removal and Sample Recovery, 6-10
xii
Contents
References, E-1
List of References, E-1
xiii
Contents
Glossary
Beckman Coulter, Inc.
J Series Rotor Warranty
xiv
Illustrations
Illustrations
xv
Tables
Tables
xvi
Scope
This manual contains general information for properly preparing a rotor for centrifugation in
a Beckman Coulter J series centrifuge. This manual should be used with the individual rotor
instruction manual shipped with each rotor. The rotor manuals provide specific information for
each rotor, including special operating procedures and precautions, tube, bottle, and adapter part
numbers, and equations to calculate maximum allowable rotor speeds. Each manual has a code
number in the bottom left-hand corner of the cover page that can be used for reordering. To
reorder, call Beckman Coulter Customer Service at 1-800-742-2345 (U.S.A. or Canada), outside the
U.S., contact your local Beckman Coulter representative.
A lot of information is compiled in this manual, and we urge you to read it carefully — especially if
this is your first experience with Beckman Coulter products.
• CHAPTER 1 describes, by usage, Beckman Coulter’s currently produced J series rotors; this
should help you determine the appropriate rotor to use for a particular application. Also
included in this section is a discussion of rotor materials, components, and centrifugation
techniques.
• CHAPTER 2 describes various tubes, adapters, spacers, and cannisters to help you choose a
particular container for your application.
• CHAPTER 3 provides instructions for using tubes, bottles, cannisters, and related accessories.
• CHAPTER 4 contains step-by-step procedures for preparing a fixed angle rotor for a centrifuge
run. Similar information is available for swinging bucket rotors in CHAPTER 5, and CHAPTER 6
contains the same type of information for vertical tube and rack-type rotors. (Elutriation, zonal,
and continuous flow rotors are not covered in this manual.)
• CHAPTER 7 provides rotor, tube, and accessory care and maintenance information, as well as
some diagnostic hints. Please read it. Proper rotor care results in longer rotor life.
PN JR-IM-10AG xvii
• Several appendixes contain information that may be of special interest:
— APPENDIX A lists chemical resistances for rotor and accessory materials to help determine
compatibility with a variety of solutions.
— APPENDIX B contains Temperature Compensation Tables for various rotors.
— APPENDIX C contains reference information on some commonly used gradient materials.
— APPENDIX D provides information about separation of blood components using J series
centrifuges.
— APPENDIX E lists references for further reading.
— Glossary provides a glossary of terms.
PN JR-IM-10AG xviii
CHAPTER 1
Rotors
Introduction
This chapter is an introduction to the Beckman Coulter family of J series rotors, providing general
information on rotor design, selection, and operation. Rotor designs described are fixed angle, swinging
bucket, vertical tube, and rack type. Specific instructions for using each type of rotor are contained in
CHAPTER 4, CHAPTER 5, and CHAPTER 6. Care and maintenance information for all of these rotors is
contained in CHAPTER 7. Elutriator, continuous flow, and zonal rotors are not covered in this manual. The
elutriator rotors are described in detail in their respective rotor instruction manuals, publications JE6B-IM-
9 and JE5-IM-13; the continuous flow/zonal rotor, JCF-Z, is described in publication JCFZ-IM-12.
General Description
Rotor Designations
Beckman Coulter J series rotors are usually named according to the type of rotor and the rotor’s
maximum allowable revolutions per minute (in thousands), referred to as rated speed. For example,
the JA-12 rotor is a fixed-angle rotor with a maximum speed of 12,000 rpm. However, the naming
system for J series rotors was changed slightly in early 1994.
• Rotors released before 1994 (for example, the JA-18.1): JA designates that it is a fixed-angle rotor
used in a J series centrifuge; the 18 indicates that the rated speed of the rotor is 18,000 rpm; the
decimal unit (.1) distinguishes between different rotors with the same rated speed.
• Rotors released after January 1994 (for example, the JA-25.50): JA still designates that it is a
fixed-angle rotor used in a J series centrifuge; the 25 still identifies the rated speed of the rotor
(25,000 rpm); but the decimal unit (.50) describes the nominal volume of the largest tube or
bottle (in mL) used in the rotor.
PN JR-IM-10AG 1-1
Rotors
General Description
Containers in fixed-angle rotors (designated JA) are held at an angle to the axis of rotation in tube
cavities.
Containers in J-Lite fixed-angle rotors (designated JLA) are also held at an angle to the axis of rotation;
the rotor construction results in reduced overall weight.
Containers in swinging-bucket rotors (designated JS) are held in rotor buckets or multitube carriers
attached to the rotor body by hinge pins. The buckets or carriers swing out to a horizontal position
as the rotor accelerates.
Tubes in vertical-tube rotors (designated JV) are held parallel to the axis of rotation. These rotors
have plugs, screwed into the rotor cavities over sealed tubes, that keep the tubes in the cavities and
provide support for the hydrostatic forces generated by centrifugation.
Tubes in the rack-type rotor (designated JR) are held in gamma-counter racks. Racks are loaded into
special plastic trays, which are then loaded into carriers at a resting angle. During centrifugation,
the carriers swing out to a completely horizontal position.
1-2 PN JR-IM-10AG
Rotors
General Description 1
Materials
Most Beckman Coulter J series rotors are made of aluminum and are anodized to protect the metal
from corrosion. (The JS-13.1 and JS-7.5 rotors are painted with polyurethane paint and are not
anodized.) The anodized coating is a thin, hard layer of aluminum oxide formed electrochemically
in the final stages of rotor fabrication. A black or colored dye may be applied over the oxide for rotor
family identification. The coating can be damaged if careful cleaning procedures are not followed.
Therefore, it is especially important to clean aluminum rotors with brushes that will not scratch the
anodized coating and to use a noncorrosive, neutral-pH detergent. Refer to CHAPTER 7 for cleaning
and maintenance procedures.
PN JR-IM-10AG 1-3
Rotors
Rotor Selection
Drive Pins
Currently produced J series rotors have drive pins in the drive hole. These pins mesh with teeth on
the centrifuge drive spindle hub when the rotor is installed to ensure that the rotor does not slip on
the hub during initial acceleration. Most drive pins are oriented horizontally (or angled) in the drive
hole; however, some are oriented vertically.
Rotor Selection
Rotors used in Beckman Coulter J series centrifuges are listed in Table 1.1. General rotor
specifications for each fixed-angle rotor are in Table 4.1, swinging buckets in Table 5.1, and vertical-
tube and rack-type in Table 6.1. Detailed descriptions of each rotor are included in the applicable
rotor manual.
1-4 PN JR-IM-10AG
Table 1.1 Rotors Used in Beckman Coulter J Series Centrifugesa
PN JR-IM-10AG
Avanti JXN-26
Avanti JXN 30
Avanti J-30 I
Avanti J-HC
Avanti J-E
Nominal Max Max
(J2-MC)
(J6-MC)
(J2-HC)
(J6-HC)
(J2-HS)
Rotor Speed RCF
J6-MI
Rotor Capacity (rpm) (x g) k Factor
Fixed Angle
Rotor Selection
JA-18 1 liter 18,000 47,900 56x X X X X X X X X X X X X X X
Rotors
1-5
1
Table 1.1 Rotors Used in Beckman Coulter J Series Centrifugesa (Continued)
1-6
Rotor Selection
Rotors
(Avanti J-20 XP Series)
Avanti JXN-26
Avanti JXN 30
Avanti J-30 I
Avanti J-HC
Avanti J-E
Nominal Max Max
(J2-MC)
(J6-MC)
(J2-HC)
(J6-HC)
(J2-HS)
Rotor Speed RCF
J6-MI
Rotor Capacity (rpm) (x g) k Factor
Swinging Bucket
Avanti JXN-26
Avanti JXN 30
Avanti J-30 I
Avanti J-HC
Avanti J-E
Nominal Max Max
(J2-MC)
(J6-MC)
(J2-HC)
(J6-HC)
(J2-HS)
Rotor Speed RCF
J6-MI
Rotor Capacity (rpm) (x g) k Factor
Rotor Selection
(JV-20) 312 mL 20,000 41,619 206 X X X
Rotors
(JR-3.2) 320 mL 2,280 25,606 X X X
1-7
1
Table 1.1 Rotors Used in Beckman Coulter J Series Centrifugesa (Continued)
1-8
Rotor Selection
Rotors
(Avanti J-20 XP Series)
Avanti JXN-26
Avanti JXN 30
Avanti J-30 I
Avanti J-HC
Avanti J-E
Nominal Max Max
(J2-MC)
(J6-MC)
(J2-HC)
(J6-HC)
(J2-HS)
Rotor Speed RCF
J6-MI
Rotor Capacity (rpm) (x g) k Factor
Zonal and Continuous Flow (see applicable rotor manual for rotor description and use)
Selection of a rotor depends on a variety of factors, such as sample volume, number of sample
components to be separated, particle size, run time, required quality of separation, type of
separation, and the centrifuge in use. Fixed-angle, swinging-bucket, vertical-tube, and rack-type
rotors are designed to provide optimal separations for a variety of sample types. (For especially
large sample volumes, continuous flow and zonal rotors are available.)
PN JR-IM-10AG 1-9
Rotors
Rotor Selection
Fixed-Angle
Rotors
rmin rmax
Pathlength
Swinging-Bucket
Rotors
rmin rmax
Pathlength
Vertical-Tube
Rotors
rmin rmax
Pathlength
* Dark gray represents pelleted material, light gray is floating components, and bands are indicated by black lines.
1-10 PN JR-IM-10AG
Rotors
Rotor Selection 1
ln ( rmax ⁄ rmin ) 10 13 EQ 1
- × ------------
k = ----------------------------------
ω2 3600
where ω is the angular velocity of the rotor in radians per second (2πRPM/60, or ω = 0.10472 × rpm),
rmax is the maximum radius, and rmin is the minimum radius.
After substitution,
11 EQ 2
(2.533 × 10 ) ln ( rmax ⁄ rmin )
k = ------------------------------------------------------------------------
-
rpm 2
This factor can be used in the following equation to estimate the time t (in hours) required for
pelleting:
k EQ 3
t = ---
s
where s is the sedimentation coefficient* of the particle of interest in Svedberg units. (Because s
values in seconds are such small numbers, they are generally expressed in Svedberg units (S), where
1 S is equal to 10-13 seconds). It is usual practice to use the standard sedimentation coefficient s20,w
based on sedimentation in water at 20°C. Clearing factors can be calculated at speeds other than
maximum rated speed by use of the following formula:
2 EQ 4
k adj = k ⎛------------------------------------------------- ⎞
rated speed of rotor
⎝ actual run speed ⎠
Run times can also be calculated from data established in prior experiments when the k factor of the
previous rotor is known. For any two rotors, a and b:
t ka EQ 5
----a- = ------
tb kb
where the k factors have been adjusted for the actual run speed used.
Figure 1.3 lists sedimentation coefficients for some common biological materials. The k factors at
rated speeds for Beckman Coulter J series rotors are provided in the table of general specifications
in each rotor use section.
PN JR-IM-10AG 1-11
Rotors
Rotor Selection
Figure 1.3 Sedimentation Coefficients (in Svedberg Units) for Some Common Biological Materials
0
1
2
Cytochrome c
3
Collagen
4 Yeast tRNA
Albumin
5
Luteinizing hormone
Soluble Proteins 6
Immunoglobulin G
7
8
Aldolase
9
Nucleic Acids
Catalase 10
α2-Macroglobulin E. coli rRNA
20
Calf liver DNA
Ribosomal subunits 40 Vesicular stomatitis virus RNA
Bacteriophage T5 DNA
60 Bacteriophage T2 & T4 DNAs
Ribosomes 80 Broad bean mottle
100
Poliomyelitis
Polysomes 200 Tobacco mosaic
Equine encephalitis
400 Viruses
Mitochondria
100 000
The centrifugal force exerted at a given radius in a rotor is a function of the rotor speed. The
nomograms for J2 series and J6 series centrifuges in Figure 1.4 and Figure 1.5 allow you to determine
relative centrifugal field (RCF) for a given radius and rotor speed. In Avanti J series centrifuges, the
RCF is calculated automatically by the centrifuge software.
Run times can be shortened by using partially filled thickwall polypropylene and polycarbonate
tubes. The short pathlength means less distance for particles to travel in the portion of the tube
experiencing greatest centrifugal force, and hence shortened run times. The k factors for half-filled
tubes can be calculated by using an approximate rmax and rav in k factor EQ 1.
1-12 PN JR-IM-10AG
Rotors
Rotor Selection 1
10 000
10 000
100
5 000
90
80
5 000
70
1 000
60
500
50
200 2 000
40
100
50
40
30 1 000
30
20
25 15 750
* Align a straightedge through known values in two columns; read the figure where the straightedge intersects the third
column.
PN JR-IM-10AG 1-13
Rotors
Rotor Selection
200
1 000
2 000
180
500
160
140
200
1 000
120
100
100 50
90
500
80 20
70 10
60 5
50 200
2
Radial Distance Relative Centrifugal Field Speed
(mm) (× g) (rpm)
* Align a straightedge through known values in two columns; read the figure where the straightedge intersects the third
column.
1-14 PN JR-IM-10AG
Rotors
Rotor Selection 1
Isopycnic Separations
A sedimentation-equilibrium, or isopycnic, method separates particles on the basis of particle
buoyant density. Each component in the sample travels through the gradient until it reaches an
equilibrium position. Particle velocity due to differences in density is given in the following
expression:
d2 ( ρp – ρc ) EQ 6
v = ------------------------------- × g
18μ
where
v= sedimentation velocity (dr/dt)
d= particle diameter
ρp = particle density
ρc = solution density
μ= viscosity of liquid media
g= standard acceleration of gravity
The gradient may be preformed before the run or generated during centrifugation. For gradients
formed by centrifugation, the time it takes to form a gradient depends on the sedimentation and
diffusion coefficients of the gradient material, the pathlength, and the rotor speed. For a given
gradient material, the shorter the pathlength and the higher the rotor speed, the faster the gradient
will form. In general, the time required for gradients to reach equilibrium in swinging-bucket rotors
will be longer than in fixed-angle rotors. One way to reduce run times is to use partially filled tubes.
Refer to the applicable rotor manual to determine the maximum allowable speed and solution
density when using partially filled tubes.
PN JR-IM-10AG 1-15
Rotors
General Operating Information
• Soft spin runs, short centrifugation runs (3 to 5 minutes) at low g-forces (2000 to 3000 × g) at
ambient temperature, are used to keep small cells or platelets in suspension while the larger
cells sediment. This type of run is used to obtain platelet-rich plasma and red blood cell
concentrate from whole blood.
• Hard spin runs are longer (5 to 7 minutes), at higher g-forces (4000 to 5000 × g), at ambient
temperatures or at 4°C, and are used to separate fresh plasma from cellular components.
Soft spin and hard spin techniques are often combined. Refer to APPENDIX D for further
information about separation of blood components by centrifugation.
NOTE Specific information about filling, sealing, and capping containers, loading rotors, etc., is contained in
later sections.
Rotor Balance
If sample quantity is limited and the rotor is not balanced, do one of the following to balance the
rotor, depending on the rotor in use:
• Load the opposite rotor cavities or buckets with tubes containing a liquid of the same density as
opposing tubes.
• Layer a low-density, immiscible liquid, such as mineral oil, on top of the sample to fill opposing
tubes to the same level.
1-16 PN JR-IM-10AG
Rotors
General Operating Information 1
Rotor Tie-Down
Knob
To secure the rotor to the drive spindle hub during centrifugation, J series
rotors are equipped with devices that screw into the hub. If the rotor is
Tie-down
Assembly left in the centrifuge between runs, tighten the tie-down device before
each run.
Rotor Knob • Some new and modified rotors have dual-locking lid mechanisms. The
dual-locking lid mechanism consists of a daisy knob that secures the
Daisy lid to the rotor, and a tie-down knob that attaches the rotor to the
Knob
centrifuge drive hub. (Daisy refers to the knob shape. The grooves
between each “petal” let your fingers grip the knob firmly and provide
leverage for turning.) The daisy knob allows you to attach the lid to the
rotor before placing the rotor into the centrifuge, and to remove the
rotor from the centrifuge with the lid attached.
Rotor
Hold-Down CAUTION
Screw
Always loosen the rotor knob before loosening the daisy knob to avoid
jamming the knobs.
Torque Bar
• Other rotors are secured to the centrifuge drive spindle hub by a tie-
down screw. A torque bar is supplied with the rotor to provide leverage
to securely fasten the rotor.
* For example, two, three, four, or sic tubes can be arranged symmetrically in a six-place rotor.
PN JR-IM-10AG 1-17
Rotors
General Operating Information
Overspeed Protection
Rotors are specifically designed to withstand a maximum load (that is, volume and density of the
rotor contents) at rated speed. At greater speeds, or at rated speeds with heavier loads, rotors are
subject to failure.
• In Avanti J series centrifuges, an electronic recognition system identifies the rotor, thereby
limiting speed to the rated speed of the rotor.
• In J2 and J6 series centrifuges with analog controls, the rotor speed is limited by the physical
properties of the rotor. Friction created by the air in the centrifuge chamber interacting with
the rotor surfaces during centrifugation in most cases prevents rotors from exceeding their
rated speeds.
• In microprocessor-controlled J2 and J6 series centrifuges, internal circuitry monitors the rotor
speed and prevents a rotor from exceeding its rated speed. The rotor entry code listed in the
applicable rotor manual sets the allowable speed.
At rated speeds with heavier loads, rotors are subject to failure. It is the operator’s responsibility to
limit rotor speed when centrifuging dense solutions or when using heavy containers; refer to
Allowable Run Speeds below.
SPEED RPM/RCF
1-18 PN JR-IM-10AG
Rotors
General Operating Information 1
• Dense Solutions. When using dense solutions (> 1.2 g/mL) in J2 series rotors, determine
maximum run speed using the following square-root reduction formula:
ρ EQ 7
reduced run speed = maximum rated speed ------A-
ρB
where ρA is the maximum permissible density of the tube contents for a particular rotor (from
the rotor manual), and ρB is the actual density of the tube contents to be centrifuged.
When using dense solutions in J6 series rotors, determine maximum run speed using the
following square-root reduction formula:
A EQ 8
reduced run speed = maximum rated speed ----
B
where A is 2500 grams for JS rotors or 1500 grams for the JR-3.2 rotor, and B is the weight in
grams of a total load (bucket with adapter and sample; bucket with blood bag cup and filled
blood bag; tray with racks, tubes, and sample).
NOTE The maximum speed for Avanti J or J2 series rotors in J6 series instruments is 6000 rpm with
solutions of density no greater than 2.0 g/mL. Solutions of density greater than 2.0 g/mL should not
be centrifuged.
• Critical Speed Range. The critical speed range of a rotor is the range of speeds in which, during
acceleration, the rotor shifts so as to rotate about its center of mass. While passing through this
speed range, the rotor will usually vibrate. Do not set operating speeds that are within a rotor’s
critical speed range (as listed in the rotor manual).
• Minimum Speeds. Some buckets or carriers will not achieve their full horizontal position if the
rotor is run below minimum rotating speed. Refer to the individual rotor manual for speed
requirements.
Temperature Compensation
To ensure that the rotor reaches the required temperature during centrifugation, some
temperature compensation may be required because of the mass of these rotors. Tables listing
temperature compensation units for various rotors are contained in APPENDIX B and individual
rotor manuals. Follow the instructions below for the model of centrifuge being used.
Avanti J Centrifuges
Avanti J series centrifuges provide automatic temperature compensation. Enter the run
temperature according to the instructions in your centrifuge instruction manual. No additional
input is required.
PN JR-IM-10AG 1-19
Rotors
General Operating Information
1 Turn the SET knob on the centrifuge panel to the required sample temperature.
2 Find the compensation value in APPENDIX B (or in the applicable rotor manual) that
corresponds with the required temperature and run speed.
Set the COMP dial to that setting.
(Interpolate if intermediate values are required.)
NOTE Temperature settings for J-21 series centrifuges must be empirically determined.
1 Find the compensation value in APPENDIX B (or in the applicable rotor manual) that
corresponds with the required temperature and run speed.
(Interpolate if intermediate values are required.)
2 Turn the SET knob on the centrifuge control panel to the required sample temperature.
1-20 PN JR-IM-10AG
Rotors
General Operating Information 1
Microprocessor-Controlled Centrifuges
Operating temperatures for most rotors are contained in memory in microprocessor-controlled
centrifuges (model J6-MI, and discontinued models J6-MC, J2-MI, J2-21M, J2-MC, and J2-21M/E). Set
temperature compensation as follows for rotors not in centrifuge memory.
1 Press the TEMP key on the centrifuge control panel and then use the keypad to enter the
required sample temperature.
2 Find the compensation value in APPENDIX B (or in the applicable rotor manual) that
corresponds with the required temperature and run speed.
PN JR-IM-10AG 1-21
Rotors
General Operating Information
1-22 PN JR-IM-10AG
CHAPTER 2
Introduction
This section describes various labware used in Beckman Coulter J series rotors. General instructions for
using containers follow in CHAPTER 3. Care and maintenance instructions are in CHAPTER 7. General rotor
use instructions are in CHAPTER 4, CHAPTER 5, and CHAPTER 6. The individual rotor manual that comes
with each rotor provides specific instructions on the tubes, bottles, and accessories that can be used in a
particular rotor. *A table of chemical resistances can be found in APPENDIX A of this manual.
No single tube or bottle design or material meets all application requirements. Labware choice is
usually based on a number of factors.
• The centrifugation technique to be used, including the rotor in use, quantity of sample to be
centrifuged, need for sterilization, importance of band visibility, and so forth
• Chemical resistance—the nature of the sample and any solvent or gradient media
• Temperature and speed considerations
• Whether tubes or bottles are to be reused
Table 2.1 contains an overview of some of the characteristics of tube and bottle materials.
* A complete list of tubes, bottles, and accessories is provided in the latest edition of the Beckman Coulter High
Performance, High Speed, High Capacity Rotors, Tubes, and Accessories catalog (BR-8102), available at
www.beckmancoulter.com.
PN JR-IM-10AG 2-1
Tubes, Bottles, and Accessories
Labware Selection Criteria
Table 2.1 Characteristics and Chemical Resistances of Tube and Bottle Materialsa
Hydrocarbons (aromatic
Acids (dilute or weak)
Tube or Bottle Type
Alcohols (aliphatic)
and halogenated)
Optical Property
Acids (strong)
Puncturable
Aldehydes
Reusable
Sliceable
Ketones
Esters
Bases
Salts
thinwall transparent yes yes no S U U M S U U U U U S
polypropylene
thickwall translucent no nob yes S S S M S M M U M U S
polypropylene
Ultra-Clear transparent yes yes no S U U S U U U U U U M
polycarbonate transparent no no yes M U U M U U U U U M M
polypropylene translucent/ no nob yes S S S M S M S M M M S
transparent
polyethylene transparent/ yes no yes S S S S S S U M M M S
translucent
cellulose transparent no nob no S U U U U M S S U M S
propionate
stainless steel opaque no no yes S U S S M S S S M S M
S = satisfactory resistance M = marginal resistance U = unsatisfactory resistance
NOTE This information has been consolidated from a number of sources and is provided only as a guide to
the selection of tube or bottle materials. Soak tests at 1 g (at 20°C) established the data for most of the
materials; reactions may vary under the stress of centrifugation, or with extended contact or temperature
variations. To prevent failure and loss of valuable sample, ALWAYS TEST SOLUTIONS UNDER OPERATING
CONDITIONS BEFORE USE.
WARNING
Do not use flammable substances in or near operating centrifuges.
2-2 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Labware Types 2
Labware Types
NOTE Tubes made of cellulose nitrate were formerly popular for various separations, particularly rate-zonal
separations. Beckman Coulter discontinued the use of cellulose nitrate for tube manufacture in 1980, due
to inconsistent physical properties inherent in the material. If you currently have cellulose nitrate tubes,
dispose of them. Consult your laboratory safety officer for proper disposal procedures.
Polypropylene Tubes
Polypropylene tubes are translucent or transparent in appearance, depending on wall thickness,
and are non-wettable (although some polypropylene tubes can be chemically treated to make them
wettable). Polypropylene tubes are reusable unless deformed during centrifugation or autoclaving.
Polypropylene tubes have good tolerance to gradient media, including alkalines. They are
satisfactory for many acids, bases, alcohols, DMSO, and some organic solvents. They can be used
with or without caps in fixed-angle rotors. Speed reduction is sometimes required with these tubes
if run with less than full volume (refer to your rotor manual). Several types of polypropylene tubes
are available.
Thickwall open-top tubes offer the convenience of centrifuging partially filled tubes without
tube caps in fixed-angle and swinging-bucket rotors. Because the solution reorients during
centrifugation, the maximum partial fill volume depends on the tube angle. For greater fill volumes,
use tubes with caps. Refer to the applicable rotor manual for fill volumes and speed reduction
requirements. Thickwall polypropylene tubes are typically reusable unless deformed during
centrifugation or autoclaving.
PN JR-IM-10AG 2-3
Tubes, Bottles, and Accessories
Labware Types
Polycarbonate Tubes
Polycarbonate is tough, rigid, nonwettable, and glass-like in appearance. Polycarbonate tubes are
reusable and can be used with or without caps in fixed-angle rotors, and at least half full in
swinging-bucket rotors. Speed reduction may be required in some rotors if the tubes are not
completely filled.
Although polycarbonate tubes may be autoclaved, doing so greatly reduces the usable life of these
tubes. Cold sterilization methods are recommended. Washing with alkaline detergents can cause
failure. Crazing—the appearance of fine cracks in the tube—is the result of stress “relaxation” and
can affect tube performance. These cracks will gradually increase in size and depth, becoming more
visible. Tubes should be discarded before cracks become large enough for fluid to escape. These
tubes have good tolerance to all gradient media except alkalines (pH greater than 8). They are
satisfactory for some weak acids, but are unsatisfactory for all bases, alcohol, and other organic
solvents.
Polyethylene Tubes
Polyethylene tubes are translucent or transparent and have a good tolerance for use with strong
acids and bases. They are reusable but cannot be autoclaved. In swinging-bucket rotors, they are
used without caps, and with or without caps in fixed-angle rotors.
Ultra-Clear Tubes
Ultra-Clear tubes, made of a tough thermoplastic, are thinwall and not wettable (but can be made
wettable; see CHAPTER 3). Ultra-Clear tubes are available in two types—open-top and Quick-Seal.
They are transparent centrifuge tubes, offering easy location of visible banded samples. Standard
straight-wall Ultra-Clear tubes must be filled completely and capped for use in fixed-angle rotors.
Ultra-Clear tubes, which can be used one time only, have good resistance to most weak acids and
some weak bases, but are unsatisfactory for DMSO and most organic solvents, including all alcohols.
Ultra-Clear tubes should not be autoclaved.
2-4 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Labware Types 2
Microfuge Tubes
Microfuge tubes, 1.5-mL tubes with attached caps, are made of of clear or colored polypropylene.
The tubes are placed in adapters for use in some fixed-angle rotors. They are also used in multitube
adapters in the buckets or carriers of swinging-bucket rotors. The number and arrangement of
tubes in opposing adapters should be balanced.
Bottles
Bottles are available in polycarbonate (hard and clear), polypropylene (translucent), and Sealed
polypropylene or polycarbonate bottles, available for most fixed-angle rotors, have a three-piece
liquid-tight cap assembly. Other bottles have screw-on caps. Cap assemblies should always be
removed before autoclaving bottles. Bottle selection depends on the rotor in use and the specific
application; refer to the applicable rotor manual.
2
C
3
D
4
E
5
F
7
H
10
12
PN JR-IM-10AG 2-5
Tubes, Bottles, and Accessories
Temperature Limits
11
12 96-Cap Strip
10
9
8
7
A
B
C
D
E
F
polystyrene or polypropylene. The plates can contain up
A
B
C
G
H
to 1.2 mL per well of sample and solvent in a single 96-well
D
E
F
G
H
plate when run uncapped. When used with caps, which
come in 96-cap strips, each well accommodates 1.0 mL.
1
B
C
2
D
3
E
F
4
G
5
H
6
7
8
9
10
Temperature Limits
Each labware material has a specified temperature range. Although some high-speed centrifuges
can achieve temperatures as high as 45°C, only certain tube or bottle materials can be run under
these conditions. Most containers are made of thermoplastic materials that soften at elevated
temperatures. This temperature-induced softening, together with such factors as the centrifugal
force, the run duration, the type of rotor, previous run history, and the tube angle, can cause
labware to collapse. Therefore, if high-temperature runs—above 25°C—are required, it is best to
pretest labware under the actual experimental conditions, using buffer or gradient of similar
density rather than a valuable sample. (Stainless steel tubes can be centrifuged at any temperature.)
• Plastic labware has been centrifuge tested for use at temperatures between 2 and 25°C. For
centrifugation at other temperatures, pretest tubes under anticipated run conditions.
• If plastic containers are frozen before use, make sure that they are thawed to at least 2°C prior
to centrifugation.
2-6 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Spacers and Floating Spacers 2
Adapters
Many rotors can accommodate a variety of tube sizes by using adapters that line the tube cavity or
bucket. Adapters are fabricated of several different kinds of materials, depending on the rotor and
the tube to be used in them. Some of the common materials are acetal, modified polyphenylene
oxide, polyethermide (PEI), polyethylene, rubber, polypropylene, and glass-filled or foamed
polypropylene.
Tubes or bottles used with adapters can be filled (and capped, if applicable), according to the type
of tube and the design of the rotor being used.
Bottle Adapters
Bottles are often supported during centrifugation in bottle adapters that fit
inside the rotor buckets or cavities. The adapters are usually ribbed for strength
Bottle
and support a variety of bottle sizes.
Multitube Adapters
Adapters are used to enable centrifugation of multiple tubes in the bucket of a swinging-bucket
rotor or in a fixed-angle tube cavity.
PN JR-IM-10AG 2-7
Tubes, Bottles, and Accessories
Adapters
Bails
These adapters can also be used as tube racks in the
laboratory. The adapter disks are color-coded by the
tube size they accommodate; the number of disks used
in an adapter assembly depends upon the length of
tubes used. Refer to the applicable rotor manual to
determine the kind of adapter required for the tubes
you are using. A tube decanter is available to hold tubes
JS-4.3
securely in some adapters, allowing all tubes to be
decanted at once.
2-8 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Bottle and Tube Caps 2
The need for caps depends on such factors as the kind of rotor being used, the type of container, and
the amount of sample being centrifuged.
Some tubes must be capped before centrifugation, as in the case of thinwall tubes. The thickness
and strength of some containers, such as thickwall plastic and stainless steel tubes used in fixed-
angle and swinging-bucket rotors, allows them to be run without caps, but they must be only
partially filled. (Refer to the applicable rotor manual for allowable capless fill levels.) When greater
fill volumes are required in these tubes, caps must be used for sample retention.
• Cap assemblies—threaded caps with inserts and O-rings, or one-piece caps with O-rings, that
provide a leakproof closure to accommodate a capacity container load (that is, to the bottom of
the insert).
• Threaded caps without inserts or O-rings—these are not as liquid-tight as the cap assemblies;
therefore, the meniscus must be kept lower to prevent leakage.
• Snap-on caps—these caps are simple to use but are not as liquid-tight as the cap assemblies or
threaded caps. They require an even lower meniscus to prevent leakage.
Aerosolve Canisters
Lid
The canister and lid are made of polyphenylsulfone, tube racks are made of
polypropylene, and the O-ring is ethylene-propylene rubber. Refer to
APPENDIX A, Chemical Resistances, to determine compatibilities with specific
Aerosolve chemicals.
Canister
Each canister can hold a variety of tube sizes in tube racks that are specifically
designed to fit in the canisters. The canister can also be used as a 500-mL wide-
mouth bottle.
WARNING
When centrifuging hazardous materials, always open canisters in an
Aerosolve appropriate blood or biological safety cabinet.
Tube Rack
PN JR-IM-10AG 2-9
Tubes, Bottles, and Accessories
Blood Bag Cups
Plug
Air-Vent
Filter Cup Cover
Spout
Cup
The HarvestLine System for the JLA-8.1000 and JLA-9.1000 rotors provides
a convenient method of loading, recovering, and storing samples run in
these rotors. Up to six rotor bottles are placed in the filling rack, and a
liner is placed into each bottle. The liners are loaded with sample through
a funnel or fermentor hose. The valve in the neck of each liner is then
sealed and the liner necks folded to fit inside the bottles. The bottles are
sealed with rotor plugs and cap/closures, and the sealed bottles are placed
into the rotor canisters for centrifugation. After centrifugation, the liner
valves are cut off and the supernatant decanted, either for storage or
disposal. The liners can then be heat-sealed for pellet storage or disposal.
2-10 PN JR-IM-10AG
CHAPTER 3
Introduction
This section contains general instructions for filling and capping the labware used in Beckman Coulter
J series rotors, for selecting and using the appropriate accessories, and for recovering samples after a run.
Individual rotor manuals provide specific instructions on tubes, bottles, and accessories that can be used
in a particular rotor.*
Rotor use instructions are in CHAPTER 4 for fixed-angle rotors, in CHAPTER 5 for swinging-bucket rotors,
and in CHAPTER 6 for vertical-tube and rack-type rotors. A table of chemical resistances is in APPENDIX A
of this manual. Reference information on some commonly used gradient materials is in APPENDIX C.
Gradient Preparation
Many commercial gradient formers are available. These devices usually load a tube by allowing the
gradient solutions to run down the side of the tube. The heaviest concentration is loaded first,
followed by successively lighter concentrations. This method is acceptable for wettable tubes;
however, loading a nonwettable tube (such as Ultra-Clear, polypropylene, and polycarbonate) by
allowing solutions to run down the side of the tube can cause mixing.
Added
5%
First Gradients in nonwettable tubes can be prepared using a gradient
10%
former by placing a long syringe needle or tubing to the tube bottom
and reversing the gradient chambers. In that way the lightest gradient
15% concentration is loaded first, underlayed by increasingly heavier
concentrations.
Added
Last 20%
* A complete list of tubes, bottles, and adapters is provided in the latest edition of the Beckman Coulter High Performance, High
Speed, High Capacity Rotors, Tubes & Accessories catalog (BR-8102), available at www.beckmancoulter.com.
PN JR-IM-10AG 3-1
Tubes, Bottles, and Accessories
General Filling and Sealing or Capping Requirements
See Table 3.1 for general filling and sealing or capping requirements for tubes and bottles used in
J series rotors. Maximum fill volume includes sample and gradient. Refer to individual rotor
manuals for specific filling and capping requirements.
3-2 PN JR-IM-10AG
Tubes, Bottles, and Accessories
General Filling and Sealing or Capping Requirements 3
Table 3.1 General Filling and Sealing Requirements for Tubes and Bottles
Vertical-Tube
Tube or Bottle Swinging-Bucket Rotors Fixed-Angle Rotors Rotors
Polypropylene
thinwall tubes
within 2 to 3 mm of top full with cap not used
thickwall tubes
at least 1/2 full 1/2full to max capless level or full not used
with cap
Quick-Seal tubes
full and heat sealed full and heat sealed full and heat sealed
bottles 1/2
min to max (see rotor manual) full to max (see rotor manual) not used
with screw-on cap or cap with screw-on cap or cap assembly
assembly
Ultra-Clear
open-top tubes within 2 to 3 mm of top full with cap not used
Quick-Seal tubes not used full and heat sealed full and heat sealed
Polycarbonate
thickwall tubes at least 1/2 full 1/2full to max capless level or full not used
with cap
bottles at least 1/2 full min to max (see rotor manual) with not used
screw-on cap or cap assembly
Stainless Steel
tubes any level any level with cap or cap assembly not used
Polypropylene
tubes and bottles at least 1/2 full 1/2full to max capless level or full not used
with cap or cap assembly
Polyethylene
tubes at least 1/2 full 1/2full to max capless level or full not used
with cap
Cellulose
Propionate
at least 1/2 full 1/2 full to max capless level not used
tubes and bottles
PN JR-IM-10AG 3-3
Tubes, Bottles, and Accessories
General Filling and Sealing or Capping Requirements
WARNING
Handle body fluids with care because they can transmit disease. No known test
offers complete assurance that they are free of micro-organisms. Some of the
most virulent—Hepatitis (B and C) and HIV (I–V) viruses, atypical mycobacteria,
and certain systemic fungi—further emphasize the need for aerosol protection.
Handle other infectious samples according to good laboratory procedures and
methods to prevent spread of disease. Because spills may generate aerosols,
observe proper safety precautions for aerosol containment. Do not run toxic,
pathogenic, or radioactive materials in this rotor without taking appropriate
safety precautions. Biosafe containment should be used when Risk Group II
materials (as identified in the World Health Organization Laboratory Biosafety
Manual) are handled; materials of a higher group require more than one level of
protection.
When working with potentially hazardous materials, always fill or open containers in an
appropriate hood or biological safety cabinet. Three levels of containment are offered by
Beckman Coulter, and may be used singly or combined, depending upon your application.
1. Capped tubes or bottles are designed to provide fluid containment. We strongly recommend
that all containers carrying physiological fluids be capped to prevent leakage.
2. Rotor or bucket covers are designed to minimize the possibility of fluid leakage during
centrifugation.
• Bucket covers for swinging bucket rotors help to contain fluids within the bucket in the
event of tube breakage or blood-bag failure.
• Some fixed-angle rotors have available dual-locking lid mechanisms that provide added
biosafety by allowing the rotor to be loaded into and removed from the centrifuge with the
lid in place. The rotor may be placed under a safety hood before the lid is attached or
removed.
3. Aerosolve canisters are designed to minimize the possibility of aerosol (and fluid) leakage
during centrifugation.
3-4 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Filling Open-Top Tubes 3
Swinging-Bucket Rotors
Fixed-Angle Rotors
Polycarbonate
Thickwall polycarbonate tubes can be centrifuged partially filled. Observe maximum rotor speeds
and fill volumes listed in the applicable rotor manual.
Ultra-Clear
For swinging-bucket rotors, fill to within 2 or 3 mm of the top of the tube. Refer to the applicable
rotor manual.
PN JR-IM-10AG 3-5
Tubes, Bottles, and Accessories
Capping Tubes
Polypropylene
Fill all opposing tubes to the same level.
Polyethylene
For swinging bucket and fixed-angle rotors, fill these tubes from half full to maximum level. Refer to
the applicable rotor manual.
Stainless Steel
Because of their strength, stainless steel tubes can be centrifuged while filled to any level (with all
opposing tubes filled to the same level). However, run speeds must be reduced due to their weight.
The criteria for speed reduction depends on the tube-cap material and the strength of the rotor
being used. Refer to the applicable rotor manual or Run Speeds for Stainless Steel Tubes (publication
L5-TB-072) for correct run speeds.
Capping Tubes
Caps must be used with thinwall polypropylene and Ultra-Clear tubes in fixed-angle rotors. To
prevent spillage, thickwall polypropylene , polycarbonate, and stainless steel tubes must be capped
when fill levels exceed the maximum level for uncapped tubes as listed in the applicable rotor
manual.
Cap requirements depend on the tube or bottle material, diameter, and wall thickness, as well as on the
rotor. The applicable rotor manual specifies which cap should be used with a particular tube or bottle; use
of the wrong cap could cause a rotor mishap.
• Cap assemblies—threaded caps with inserts and O-rings that provide a leakproof closure to
accommodate a capacity container load (that is, to the bottom of the insert). Single-piece cap
assemblies have the insert permanently attached.
• Threaded caps without inserts or O-rings—these are not as liquid-tight as the cap assemblies;
therefore, the meniscus must be kept lower to prevent leakage. Speed reductions may also be
required with lower fill volumes.
• Snap-on caps—these caps are simple to use but are not as liquid-tight as the cap assemblies or
threaded caps. They require an even lower meniscus to prevent leakage.
3-6 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Filling and Capping Tubes 3
To prevent spillage and provide support, polycarbonate and polypropylene bottles used in fixed-
angle rotors must be capped when fill levels exceed the maximum level allowed for uncapped
bottles. Bottles should be filled to maximum fill levels when spun at full rated speeds. Unless
specified otherwise, the minimum recommended volume for bottles is half full; this will require
reduced rotor speed for optimum labware performance. Refer to the applicable rotor manual for
bottle fill levels and cap requirements.
Three-Piece Assemblies
Cap bottles with three-piece cap assemblies as follows:
1 Be sure the O-ring, plug, and bottle lip are dry and free of lubrication.
3 Insert the plug into the neck of the bottle, ensuring that no fluid contacts the O-ring.
Cap
Plug
O-ring
Bottle
Lip
Scribe Mark
PN JR-IM-10AG 3-7
Tubes, Bottles, and Accessories
Filling and Loading Cups in the JS-5.0 Rotor
NOTE Four labware cups must be used for every run and must be balanced to within 25 grams of
each other. Do not load the rotor with two filled cups and two empty cups.
1 Insert four labware cups into two cup racks with the cup latch hinges toward the center of
the racks.
2 Make sure that the gaskets and sealing surfaces on each cup and cover are clean and dry.
a. Place a gasket around the top edge of each cup, carefully pushing the gasket down until it
is fully seated on the cup.
b. Use green gaskets (369261) if you are using liners.
c. Use red gaskets (369257) if you are using cups alone, with or without partitions.
4 You may fill the cups now, or close the lid and fill through the spout.
NOTE If liners are not used, partitions (369259) may be inserted into the slots inside the cups.
Remove the red cup gaskets (369257) before inserting partitions, and be sure to reinstall
the gaskets.
3-8 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Filling and Sealing Quick-Seal Tubes 3
6 If the cups were not filled previously, load sample into each cup through the cover spout using
a funnel, tubing (1.27-cm [1/2-in.] O.D.), or a pipette.
a. Use the fill line indicators to assist in filling all four cups to the same level.
All four cups must balance to within 25 grams of each other.
b. When loading is complete, snap a plug into place in each cover spout.
Fill each tube to the base of the neck, using a syringe with a 13-gauge or smaller needle.* A small air
space (no larger than 3 mm) may be left, but an air bubble that is too large can cause the tube to
deform, disrupting gradients or sample. Spacer and/or floating spacer requirements for Quick-Seal
tubes are described in the individual rotor manuals. The neck of the tube should be clean and dry
before sealing.
There are two tube sealers for use with Quick-Seal tubes—the hand-held Cordless Tube Topper, and
the older tabletop model (no longer available). Refer to How to Use Quick-Seal Tubes with the
Beckman Coulter Cordless Tube Topper (publication IN-181) for detailed information about the Tube
Topper. Instructions for using the older tabletop tube sealer are in How to Use Quick-Seal Tubes with
the Beckman Tube Sealer (publication IN-163).
Quick-Seal tubes are heat-sealed quickly and easily using the Beckman Coulter Cordless Tube
Topper (see Figure 3.1). The following procedures provide the two methods for heat-sealing Quick-
Seal tubes using the hand-held Tube Topper. Use the applicable tube rack listed in the rotor manual.
* A sample application block (part number 342694) is available for holding and compressing tubes, and can be used to
layer samples on preformed gradients in polypropylene Quick-Seal tubes.
PN JR-IM-10AG 3-9
Tubes, Bottles, and Accessories
Filling and Sealing Quick-Seal Tubes
Charging Stand
Pushbutton
Tip
CAUTION
Before plugging in the Tube Topper, be sure that you have a proper power source
(120 V, 50 or 60 Hz). Charge your Cordless Tube Topper only in the charging stand
supplied with it.
WARNING
Touching the heated tip of the Tube Topper will cause burns. When the pushbutton
is pressed, the tip heats almost immediately. Make sure the pushbutton is turned
to LOCK position unless you are actually sealing a tube.
2 Place a seal former on each tube stem. (The polytetrafluorethylene coating on the seal formers
is permanent.
Do not scratch the interior of the formers, as you may damage this coating.)
Seal Former
3-10 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Filling and Sealing Quick-Seal Tubes 3
3 Seal each tube using Method A (With the Seal Guide) or Method B (Without the Seal Guide).
Method A is preferable when sealing smaller tubes or when resealing a tube that leaks.
CAUTION
Always keep the Tube Topper in its charging stand when not in use. Do not lay the
unit against any surface after use until the tip has cooled (3 to 5 minutes after
shut off).
Seal Guide
1.
a. Place a seal guide (with the flat side down) over the seal
former.
PN JR-IM-10AG 3-11
Tubes, Bottles, and Accessories
Filling and Sealing Quick-Seal Tubes
Heat Sink
Small
End
e. Place the heat sink (small end) over the cap for 2 to 3
seconds while the plastic cools—do NOT let the seal former
Removal Tool
pop up. (If the seal former does pop up, the tube may not
have an adequate seal and may need to be resealed.)
f. Remove the heat sink and seal guide. When the seal former
cools, remove it by hand or with the removal tool (361668).
Save the seal guide and former for future use.
NOTE Always apply the tip of the Tube Topper vertically to the
seal former. Apply gentle pressure when sealing the tube.
1.
a. Turn the Tube Topper pushbutton to USE position. Press
the pushbutton and wait 3 to 5 seconds for the tip to heat.
3-12 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Filling and Sealing Quick-Seal Tubes 3
Immediately
4 After completing either heat-sealing method, squeeze the tube gently (if the tube contents may
be disturbed) to test the seal for leaks. If the tube does leak, try resealing it using Method A.
5 The tube is now ready for centrifugation. Seal the remaining tubes.
PN JR-IM-10AG 3-13
Tubes, Bottles, and Accessories
Filling and Sealing Quick-Seal Tubes
Cap Strips
Available sterile or nonsterile 96-cap strips can be used with deep-well plates. (When these caps are
used, the capacity of each well is reduced to 1.0 mL.)
11
12 96-Cap Strip
10
9
8
7
6
5
4
3
2
1
A
B
C
D
E
F
G
H
A
B
C
D
E
F
G
H
Roller
(538618)
Seal &
Sample
Foil Lid
(538619)
Using Adapters
Tubes and bottles used with adapters can be filled (and capped, if applicable) according to the type
of container and the design of the rotor being used.
3-14 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Filling and Sealing Quick-Seal Tubes 3
2 Place the base and bail in an empty bucket or on the lab bench (not in the rotor).
3 Position one of the disks so that its grooves are aligned with the bail.
Push the disk down until the bail snaps into the grooves.
4 Add more disks until the height of the assembly is nearly as tall as the tubes you will be using.
(If the height of the disks is very tall, you may have to push the bail into the grooves of the top
disks by hand.)
Remove or add disks to the bail to accommodate shorter or longer tubes.
If the tubes fit too snugly in the adapter’s rubber base, apply a light film of dusting power, such
as talcum powder, to prevent the tubes from sticking.
Bails
Disks
Base
PN JR-IM-10AG 3-15
Tubes, Bottles, and Accessories
Using Aerosolve Canisters
Aerosolve canisters can be used in the JS-4.3 rotor to minimize aerosol leakage and liquid spills from
rotor buckets during centrifugation. Each canister can hold a variety of tube sizes in tube racks that
are specifically designed to fit in the canisters. The canister can also be used as a 500-mL wide-
mouth bottle.
WARNING
When centrifuging hazardous materials, always open canisters in an appropriate
hood or biological safety cabinet.
Lid
O-Ring
Aerosolve
Cannister
2 Before placing the canister in a bucket, remove the bucket-cover O-ring seated on the ledge
inside the bucket.
If this O-ring is not removed, a vacuum will be created between the bucket and canister that will
make removing the canister from the bucket difficult.
Remove
O-ring
NOTE Do not run chloroformed samples in Aerosolve canisters. Chloroform vapors can damage
the canister material.
3-16 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Using Aerosolve Canisters 3
3 Fill the canister as described under Using Canisters as Wide-Mouth Bottles, or Using Canisters with
Tube Racks, below.
4 Screw the lid on until closing resistance is first felt, then tighten it an additional 60 degrees.
The scribe marks around the rim of the canister and the corrugated finger grips on the lid are
all placed 60 degrees apart.
B A
60°
Finger
Scribe Grips
Marks
1 Fill each canister only to the fill-level line (maximum is 500 mL of 1.2 g/mL liquid).
Fill
Level
Line
2 Run another canister, filled to the same level with liquid of the same density, in the opposite
bucket.
PN JR-IM-10AG 3-17
Tubes, Bottles, and Accessories
Using Aerosolve Canisters
1 Press a rubber cushion (if applicable—see Table 3.2) into each tube hole in the rack base.
Aerosolve
Tube Rack
NOTE Partially filled tube racks should contain the same number of balanced tubes. Each
tube in a rack must be balanced by a tube in a diametrically opposed position in the
opposite rack.
3-18 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Using Blood Bag Cups 3
WARNING
Ask your laboratory safety officer to advise you about the level of containment
required for your application and about the proper decontamination or
sterilization procedures to follow if fluids escape from containers.
Different cups, color-coded for capacity identification, can accommodate single, double, triple, or
quad pack blood bags. Refer to the applicable rotor manual to determine the correct blood bag cup
to use. Do not pour liquid directly into blood bag cups. Fit blood bags into cups before loading the cups into
the rotor buckets. Stuffing blood bags directly into the rotor while it is installed in the centrifuge can
trip the imbalance detector.
PN JR-IM-10AG 3-19
Tubes, Bottles, and Accessories
Sample Recovery
2 Sandwich the tubing between the blood bag and any transfer packs.
Blood
Bag
Label
3 Make sure the loaded blood bag cups opposite each other on the rotor yoke are approximately
the same weight (within 1 gram).
(Balancing pads can be used with some rotors, if necessary, to maintain weight balance.)
Load blood bag cups into the rotor buckets. To reduce the possibility of bag breakage, align the
blood bag seam with the rotor pivot pins with the label facing out (away from the axis of rotation).
Sample Recovery
CAUTION
If disassembly reveals evidence of leakage, you should assume that some fluid
escaped the container or rotor. Apply appropriate decontamination procedures to
the centrifuge, rotor, and accessories.
You can recover labware from most J series rotors while the rotor or yoke remains in the centrifuge.
• Rotor buckets or carriers can be removed from the rotor yoke, then unloaded on a lab bench or
table or under a protective hood. Blood bags must always be removed from blood bag cups
outside of the centrifuge.
• You can remove the lid from most fixed-angle rotors and extract the tubes or bottles using a
removal tool (specified in the applicable rotor manual).
NOTE Vertical-tube rotors cannot be unloaded inside the centrifuge. The rotor must be removed
from the centrifuge and placed in a rotor vise to loosen the tube cavity plugs.
Sample recovery depends on the type of labware used, the component(s) isolated, and the analysis
desired. The Beckman Universal Fraction Recovery System (343890) can be useful when recovering
sample from tubes (see publication L5-TB-081).
3-20 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Sample Recovery 3
Capped Tubes
The usual methods of recovering supernatants or pellets include decanting or withdrawing the
gradient and scraping pellets from the tube bottom.
2 If tubes will be reused, scrape pellets out with a plastic or wooden tool; scratches on tube
interiors caused by abrasive or sharply pointed tools can result in tube failure during
subsequent runs.
JS-5.0 Cups*
1 Remove the rotor lid and hang it on the black rubber block on the inside of the centrifuge door.
2 Remove the plug from the labware cup cover and pour the supernatant out of the cup through
the spout.
Or, remove the cup cover and pour the supernatant over the cup edge.
4 If a liner was not used, first remove the red cup gasket (369257), remove the partition (if used),
and then use the spatula (367891) to remove pellet from the cup.
Do not use a metal tool to remove pellet, as metal could damage the cup and shorten its
useful life.
367891
* Beckman Coulter recommends Cole-Parmer heat sealer Model U-03018-10, adjusted to setting 3 or 4. Contact Cole-
Parmer at (800) 323-4340, by Fax at (847) 247-2929, or at www.coleparmer.com.
PN JR-IM-10AG 3-21
Tubes, Bottles, and Accessories
Sample Recovery
Quick-Seal Tubes
There are several methods of recovering fractions from Quick-Seal tubes. One of the following
procedures may be used.
Cut Quick-Seal NOTE If you plan to collect particles from the tube side or
stem here to
provide an
bottom, first create an air passage by snipping the
air inlet stem or inserting a hollow hypodermic needle in the
top of the tube.
• Puncture the side of the tube just below the band with
a needle and syringe and draw the sample off. Take
care when piercing the tube to avoid pushing the
needle out the opposite side.
Sample out
For additional information on fraction recovery systems available from Beckman Coulter, refer
to the latest edition of High Performance, High Speed, High Capacity Rotors, Tubes & Accessories
(publication BR-8102) available at www.beckmancoulter.com.
3-22 PN JR-IM-10AG
Tubes, Bottles, and Accessories
Making Ultra-Clear Tubes Wettable 3
The following method of making Ultra-Clear tubes wettable has proven successful for some users:
1. Polyvinyl alcohol (2 g) was dissolved in distilled water (50 mL) by stirring and heating to gentle
reflux.
2. Isopropanol (50 mL) was slowly added to the hot solution and stirring and heating continued
until a clear solution was obtained.
3. The solution was then allowed to cool to room temperature.
4. Ultra-Clear tubes were filled with the coating solution, then aspirated out with a water pump
after 15 minutes, leaving a thin film on the tube walls. A small amount of solution that collected
in the tube bottoms after standing was removed with a pipette.
5. The tubes were left open to dry at room temperature overnight, then filled with distilled water.
After standing overnight at room temperature, the distilled water was poured out.
6. Finally, the tubes were briefly flushed with water, tapped to remove excess liquid, and left to
dry.
PN JR-IM-10AG 3-23
Tubes, Bottles, and Accessories
Making Ultra-Clear Tubes Wettable
3-24 PN JR-IM-10AG
CHAPTER 4
Introduction
This section contains instructions for using fixed-angle rotors in J series centrifuges. In addition to these
instructions, observe procedures and precautions provided in the applicable rotor and centrifuge manuals.
Refer to CHAPTER 2 for labware selection information, and CHAPTER 3 for recommended filling and
sealing or capping requirements and for sample recovery procedures. Refer to CHAPTER 7 for information
on the care of rotors and accessories.
Description
Fixed-angle rotors (see Figure 4.1) are general-purpose rotors that are especially
useful for pelleting subcellular particles and in short-column banding of viruses and
subcellular organelles. Refer to Table 4.1 for general rotor specifications.
Tubes in fixed-angle rotors are held at an angle (usually 20 to 45 degrees) to the axis of
rotation. The tube angle shortens the particle pathlength compared to swinging-
bucket rotors, resulting in reduced run times. Tubes can be placed directly in a rotor
cavity if the diameters of the tube and the cavity are the same. Using adapters, more
than one type and size of tube can be centrifuged together, provided that the load is
properly balanced.
O-rings, made of Buna N rubber, are located in the rotor lid. The O-rings help to
maintain atmospheric pressure inside a fixed-angle rotor during centrifugation, when
they are properly lubricated.
A tie-down device or lid-locking knob is used to secure the rotor to the centrifuge
drive spindle hub before the run begins.
PN JR-IM-10AG 4-1
Using Fixed-Angle Rotors
Description
rmin
rav
rmax
25°
rmin
rav
rmax
Axis of Rotation
JA-25.15
20°
rmin
rav
rmax
Axis of Rotation JLA-10.500
4-2 PN JR-IM-10AG
Using Fixed-Angle Rotors
Description 4
Table 4.1 General Specifications for Beckman Coulter J Series Fixed-Angle Rotors
PN JR-IM-10AG 4-3
Using Fixed-Angle Rotors
Description
Table 4.1 General Specifications for Beckman Coulter J Series Fixed-Angle Rotors (Continued)
4-4 PN JR-IM-10AG
Using Fixed-Angle Rotors
Description 4
Table 4.1 General Specifications for Beckman Coulter J Series Fixed-Angle Rotors (Continued)
PN JR-IM-10AG 4-5
Using Fixed-Angle Rotors
Description
CAUTION
Always loosen the rotor knob before loosening the daisy knob to avoid jamming
the knobs.
O-Ring
Sealing Surface A feature of many Beckman Coulter fixed-angle rotors is a patented
fluid-containment annulus, located below the O-ring sealing
Fluid surface. If tubes are overfilled or if leakage occurs during
Containment
Annulus
centrifugation, the annulus holds enough volume that all of the
liquid is kept inside the rotor—even if all tubes leak at the same
time. This feature virtually eliminates the escape of liquid into the
centrifuge chamber.
NOTE Although rotor components and accessories made by other manufacturers may fit in the
Beckman Coulter rotor you are using, their safety in the rotor cannot be ascertained by Beckman Coulter.
Use of other manufacturers’ components or accessories in the Beckman Coulter rotor may void the rotor
warranty, and should be prohibited by your laboratory safety officer. Only the components and
accessories listed in the applicable rotor manual should be used.
NOTE JLA-8.1000 and JLA-9.1000 rotors run only the specially designed bottles with polypropylene AutoVent
plug and polyphenylsulfone (PPSU) or polyphenylene sulfide (PPS) cap/closures. Refer to the applicable
rotor manual for instructions on use of these bottles and accessories.
4-6 PN JR-IM-10AG
Using Fixed-Angle Rotors
Rotor Preparation and Loading 4
For runs at other than room temperature, refrigerate or warm the rotor beforehand for fast equilibration.
1 Make sure that the rotor and lid are clean and show no signs of corrosion or cracking.
3 Verify that the tubes and bottles being used are listed in the applicable rotor manual.
Rotor Preparation
1 Be sure that metal threads in the rotor are clean and lightly but evenly lubricated with Spinkote
lubricant (306812).
Also ensure that O-rings are lightly but evenly coated with silicone vacuum grease (335148).
PN JR-IM-10AG 4-7
Using Fixed-Angle Rotors
Operation
Operation
Refer to the applicable centrifuge instruction manual for detailed operating information. For low-
temperature runs, precool the rotor in the centrifuge or in a refrigerator before use—especially
before short runs—to ensure that the rotor reaches the set temperature. (To ensure that the rotor
reaches the required temperature during centrifugation, some temperature compensation may be
required because of the mass of these rotors. Refer to APPENDIX B or to the applicable rotor manual
for tables listing temperature compensation units for various rotors.)
ROTOR • If you are using an Avanti J series centrifuge (except J-E), select
SPEED Rotor :
Speed :
JA-25.50
0 RPM
the rotor number.
Time : 00:00
TIME Temp : 25°C
TEMP
Accel : MAX • If you are using an Avanti J-E or a microprocessor-controlled J2
A/D
or J6 series centrifuge, enter the rotor code (if the JA-10 rotor is
OPT
used for example, enter code 10).
4-8 PN JR-IM-10AG
Using Fixed-Angle Rotors
Operation 4
CAUTION
The centrifuge drive spindle can be bent or broken if the rotor is forced sideways
or dropped onto it. Install the rotor by centering it over the spindle and carefully
lowering it straight down.
1 Carefully lower the rotor straight down onto the drive spindle.
a. Rotate it by hand until the drive pins seat on the drive spindle hub.
• In older model centrifuges—be sure the pins in the rotor drive hole are located in the
grooves of the drive spindle hub.
Rotor
Drive Pins (2)
Hub
Grooves (4) Hub
Drive
Spindle
Assembly
• In newer model centrifuges—be sure the pins in the rotor drive hole are not sitting on
top of the teeth on the drive spindle hub.
Teeth (2)
Hub
Drive
Spindle
Assembly
CAUTION
The pins located in the rotor drive hole must be seated correctly on the centrifuge
drive spindle. Running a rotor that is not seated properly may result in severe
rotor damage.
2 After the rotor is seated on the drive spindle hub, place the lid on the rotor.
PN JR-IM-10AG 4-9
Using Fixed-Angle Rotors
Operation
NOTE The JA-18 rotor must be run with the lid on in Avanti J series centrifuges.
CAUTION
If disassembly reveals evidence of leakage, you should assume that some fluid
escaped the container or rotor. Apply appropriate decontamination procedures to
the centrifuge, rotor, and accessories.
1 Unscrew the rotor lid knob to release the rotor from the spindle hub.
NOTE Labware can be recovered from most fixed-angle rotors while the rotor remains in the centrifuge.
You can remove the lid and extract the tubes or bottles using the removal tool specified in the
applicable rotor manual. If the rotor is left in the centrifuge between runs, be sure that it is securely
tied down before each run. Remove the rotor regularly and clean the drive spindle assembly.
4-10 PN JR-IM-10AG
Using Fixed-Angle Rotors
Operation 4
2 To remove the rotor, lift it straight up and off the drive spindle.
NOTE If the rotor sticks to the drive spindle, screw the short end of the rotor lifting handle
assembly into the threaded opening to force the rotor off of the drive spindle hub.
Lubrication of the centrifuge drive spindle hub with Spinkote should prevent the rotor from
sticking on all centrifuges except Avanti J series. Avanti J series centrifuges have acetal rings
on the spindle hubs to prevent sticking and do not require lubrication.
Lifting
Handle
Assembly
(346965)
3 Remove spacers, tubes, and bottles with the appropriate removal tool.
Quick-Seal Tube
Removal Tool
(361668)
PN JR-IM-10AG 4-11
Using Fixed-Angle Rotors
Operation
4-12 PN JR-IM-10AG
CHAPTER 5
Introduction
This section contains instructions for using swinging-bucket rotors in J series centrifuges. In addition to
these instructions, observe procedures and precautions provided in the applicable rotor and centrifuge
manuals.
Refer to CHAPTER 2 for tube selection information, and CHAPTER 3 for recommended labware filling and
sealing requirements and for sample recovery procedures. Refer to CHAPTER 7 for information on the care
of rotors and accessories.
Description
Swinging-bucket rotors (see Figure 5.1) are normally used for density gradient
separations, either isopycnic or rate zonal. Refer to Table 5.1 for general rotor
specifications. A tie-down device or lid-locking knob is used to lock the rotor to the
centrifuge drive hub before the run begins.
Tubes or bottles in swinging-bucket rotors are held in the rotor buckets that are
attached to the rotor body by hinge pins. The buckets swing out to horizontal
position as the rotor is accelerated, and stay horizontal until rotor deceleration
begins. During deceleration, the buckets gradually return to vertical position.
NOTE Although rotor components and accessories made by other manufacturers may
fit in the Beckman Coulter rotor you are using, their safety in the rotor cannot be
ascertained by Beckman Coulter. Use of other manufacturers’ components or
accessories in the rotor may void the rotor warranty, and should be prohibited by your
laboratory safety officer. Only the components and accessories listed in the applicable
rotor manual should be used.
PN JR-IM-10AG 5-1
Using Swinging-Bucket Rotors
Description
rmin
rav
rmax JS-7.5
Axis of Rotation
rmin
rav
rmax
Axis of Rotation
JS-4.3
rmax
rav
rmin
JS-13.1
Axis of Rotation
5-2 PN JR-IM-10AG
Using Swinging-Bucket Rotors
Description 5
Table 5.1 General Specifications for Beckman Coulter J Series Swinging-Bucket Rotors
PN JR-IM-10AG 5-3
Using Swinging-Bucket Rotors
Description
Table 5.1 General Specifications for Beckman Coulter J Series Swinging-Bucket Rotors (Continued)
Labware
Swinging-bucket rotors can accommodate a variety of tubes, bottles, multiwell titer plates,
and blood bags, listed in individual rotor manuals. Refer to CHAPTER 3 for tube filling and
sealing requirements. Observe the maximum rotor speeds and fill volumes listed in the applicable
rotor manual.
5-4 PN JR-IM-10AG
Using Swinging-Bucket Rotors
Rotor Preparation and Loading 5
For runs at other than room temperature, refrigerate or warm the rotor beforehand for fast equilibration.
1 Make sure that the rotor and buckets or carriers are clean and show no signs of corrosion or
cracking.
3 Verify that the tubes, bottles, or carriers being used are listed in the applicable rotor manual.
4 If fluid containment is required, use capped tubes or bottles and/or Aerosolve canisters.
Beckman Coulter strongly recommends capping all containers carrying physiological fluids to
prevent leakage.
Rotor Preparation
1 Be sure that metal threads in the rotor yoke are clean and lightly but evenly lubricated with
Spinkote lubricant (306812).
2 Ensure that O-rings are in good condition and are lightly but evenly coated with silicone
vacuum grease (335148).
3 Ensure that all sealing surfaces are smooth and undamaged for proper sealing.
PN JR-IM-10AG 5-5
Using Swinging-Bucket Rotors
Rotor Preparation and Loading
4 Before each use of the rotor, make sure that bucket pin sockets are lubricated with Paint On
Graphite Lubricant (977212) as described in CHAPTER 7.
Lubrication Points
Rotor Yoke (both pin sockets)
2 Ensure that bucket O-rings are lightly but evenly coated with silicone vacuum grease.
Do not run a bucket without an O-ring, as the bucket will leak.
3 Be sure that metal threads in the bucket caps are clean and lightly but evenly lubricated with
Spinkote lubricant.
Put bucket caps on the buckets and screw them down manually.
Hanger
Cap
O-ring
Bucket
4 Hook all buckets, loaded or empty, on to the rotor, and be sure that both hooks are on the crossbar.
All six buckets must be in the same size.
Do not intermix the smaller and larger buckets in a single run.
• If fewer than six tubes are being run they must be arranged symmetrically in the rotor.
• Opposing tubes must be filled to the same level with liquid of the same density.
5-6 PN JR-IM-10AG
Using Swinging-Bucket Rotors
Rotor Preparation and Loading 5
CAUTION
The centrifuge drive spindle can be bent or broken if the rotor is forced sideways
or dropped onto it. Install the rotor by centering it over the spindle and carefully
lowering it straight down.
1 Carefully lower the rotor yoke straight down onto the drive spindle.
a. Rotate it by hand until the drive pins seat on the drive spindle hub.
Rotor
Drive Pins (2)
Hub Teeth (2)
Grooves (4) Hub
Hub
Drive Drive
Spindle Spindle
Assembly Assembly
NOTE Except for the JS-24.38 and JS-24.15 rotors, you can leave the rotor yoke in the centrifuge
between runs unless spillage has occurred—in which case you should remove the buckets or carriers
and yoke and clean the centrifuge and rotor components immediately, according to the instructions
in the centrifuge and rotor instruction manuals. The JS-24.38 and JS-24.15 rotors must be removed
from the centrifuge to install or remove buckets.
2 When the yoke is correctly seated, secure it to the drive spindle hub.
a. Rotors with tie-down knobs—hand tighten the tie-down knob. If the rotor is left in the
centrifuge between runs, tighten the knob before each run.
Tie-down Knob
b. Rotors without tie-down knobs—secure the rotor with the tie-down screw, and tighten the
screw with the torque bar or tie-down tool. If the rotor is left in the centrifuge between
runs, ensure that the screw is tight before each run.
Rotor
Tie-down
Screw
Torque Bar
PN JR-IM-10AG 5-7
Using Swinging-Bucket Rotors
Rotor Preparation and Loading
c. JS-4.2A and JS-4.2SMA—secure the rotor to the drive hub with the tie-down bolt (368518).
Tighten the bolt with the tie-down tool (368521), then remove the tool.
Tie-down Tool
(368521))
Tie-down Bolt
(368518)
NOTE Older JS-4.2A or JS-4.2SMA rotors may be secured to the drive hub with tie down bolt
(339031). Tighten the bolt with the spanner wrench (870666) or tie-down tool (368521), then
remove the wrench or tool.
d. JS-5.0—secure the rotor to the drive hub with the tie-down bolt (367824). Tighten the bolt
with the tie-down tool (368521), then remove the tool.
Rotor
Tie-down
Bolt (339031)
Spanner
Wrench
(870666)
1 To install the rotor, carefully lift it up off the rotor stand with both hands—do not lift the rotor
by the adapter—and place it on the drive hub.
Make sure that the rotor pins are perpendicular to the drive hub pins.
The pins must not rest on top of each other; turn the rotor to the right (clockwise) by hand to
check for proper installation.
2 Turn the tie-down knob to the right (clockwise) to secure the rotor.
5-8 PN JR-IM-10AG
Using Swinging-Bucket Rotors
Rotor Preparation and Loading 5
1. The buckets or carriers must be loaded symmetrically with respect to their pivotal axes (the
pivotal axis runs parallel to the crossbar, see Figure 5.2).
2. The rotor should be loaded symmetrically with respect to its center of rotation.
This means that for best results you should load opposing buckets or carriers with the same type of
labware containing the same amounts of fluid of equal density. Additionally, buckets or carriers
placed opposite each other on the rotor yoke must balance to within a certain weight, typically
10 grams (see the applicable rotor manual for details). Do not exceed the rated maximum load for
buckets or carriers.
NOTE The JS-4.2A and JS-4.2SMA swinging-bucket rotors incorporate ARIES (Automated Rotor Imbalance
Equilibrating System) “Smart Balance” technology, which provides imbalance compensation for rotors
with buckets that are up to 100 grams unbalanced due to different loading volumes or tube or bag
breakage.
Beckman Coulter supplies buckets and carriers for most rotors in weight-matched sets to make
balancing easier (the weight and date of manufacture are marked on the side of each bucket and
bottom of each carrier). Modular disk adapters are also sold in weight-matched sets. However,
there are variances in weight between sets, as well as variance in weight between previously
purchased adapters. To prevent accidental imbalance, it is important to keep matched sets of
adapters together and to weigh other adapters to be sure they are approximately the same.
Marking matched sets will help you keep them together.
PN JR-IM-10AG 5-9
Using Swinging-Bucket Rotors
Rotor Preparation and Loading
Figure 5.2 Examples of Correctly and Incorrectly Loaded Buckets and Carriers*
Balanced Load Unbalanced Load
Pivotal Axis
of Bucket
Center of
Rotation
Correct Incorrect
Pivotal Axis
Carrier
A
B
C
D
E
F
G
H
Plate
* Contents of opposing buckets must be the same and each bucket must be balanced on its pivotal axis.
5-10 PN JR-IM-10AG
Using Swinging-Bucket Rotors
Rotor Preparation and Loading 5
Loading Buckets
Buckets can be loaded before or after being installed on the rotor yoke. For best results, fill the
appropriate labware first and then load the labware into the buckets. This is especially important
when using blood bags—you can trip the imbalance detector in the centrifuge by pushing blood
bags into cups within buckets that are installed in the rotor. You can also bend the centrifuge drive
spindle.
2 Place the base and bail in an empty bucket or on the lab bench (not in the rotor).
3 Position one of the disks so that its grooves are aligned with the bail.
a. Push the disk down until the bail snaps into the grooves.
Bails
Disk
Base
4 Add more disks until the height of the assembly is nearly as tall as the tubes you will be using.
(If the height of the disks is very tall, you may have to push the bail into the grooves of the top
disks by hand.)
a. Remove or add disks to the bail to accommodate shorter or longer tubes.
b. If the tubes fit too snugly in the adapter’s rubber base, apply a light film of dusting power,
such as talcum powder, to prevent the tubes from sticking.
Place each tube in an adapter so that its weight is balanced by a tube in a diametrically opposite
position across the pivotal axis in the same adapter. Adapters placed in opposing buckets should
also be filled the same way (see Figure 5.2). If you must run only one tube in an adapter, be sure this
tube rests over the bucket’s pivotal axis.
NOTE Be sure to run a tube of the same approximate weight in the same configuration in the opposite
bucket.
PN JR-IM-10AG 5-11
Using Swinging-Bucket Rotors
Rotor Preparation and Loading
1 Load the cups so that the blood bags and tubing fit as far down as possible.
NOTE Make sure the bags stay as vertical as possible, with no folds at the top or corners. If folds are
present, blood cells could remain in the folds and then mix with the plasma when the bag is removed.
2 Sandwich the tubing between the blood bag and any transfer packs (see Figure 5.3).
3 Make sure the loaded blood bag cups opposite each other on the rotor yoke are approximately
the same weight (within limits listed in the applicable rotor manual).
In some rotors, balancing pads can be used if necessary to maintain weight balance.
CAUTION
If bucket covers or rotor lids are not used, make sure the superstructure of the
blood bag protruding from the cup does not inhibit the bucket from reaching its
horizontal position. If it does, remove the cup from the rotor and reposition the
blood bag so that it seats further into the cup. Allowing the blood bags to contact
the rotor yoke during centrifugation can cause the bucket to come off the pivot
pins and can seriously damage both the rotor and the centrifuge.
5-12 PN JR-IM-10AG
Using Swinging-Bucket Rotors
Rotor Preparation and Loading 5
Figure 5.3 Typical Blood Bag Loading Procedures (JS-24.3 Rotor Shown)
Blood Bag
Seam
Blood Bag
Label
Blood Bag
Label
Blood
Bag
Label
1 If bucket covers or rotor lids are used to help contain spills and glass particles that could result
from tube breakage, make sure cover O-rings are in good condition and lightly coated with
silicone vacuum grease.
a. Before use, inspect Aeroseal cover sealing surfaces, especially the O-ring groove.
It must be smooth and free of scratches.
Plug
(343685)
Aeroseal
Cover
O-Ring
(344660)
b. Also ensure that the top 2.54 cm (1 in.) of the bucket is clean and smooth; buckets with
scratches or gouges in this surface will not seal properly.
c. Inspect the O-ring and plug for nicks, abrasions, and other damage.
d. Replace damaged components.
PN JR-IM-10AG 5-13
Using Swinging-Bucket Rotors
Rotor Preparation and Loading
2 Load the filled buckets (and/or carriers) onto the rotor yoke pivot pins, following the
instructions in the rotor manual.
Make sure that the buckets are properly seated by gently swinging them on the pivot pins.
NOTE All positions on the rotor yoke must contain either a bucket or a microtiter plate carrier during a run.
Microplates will break if g-forces are too high. Rotor speed must be reduced when microplate
carriers are used. If microplate carriers and buckets are centrifuged in the same run, run speed must
be reduced to the speed allowable for the microplates. Refer to the applicable rotor manual for
allowable run speeds.
If only two carriers are run, they must be installed opposite each other in the rotor, and the
remaining positions on the yoke must be filled with either buckets or other carriers (they need not
be loaded) to prevent rotor imbalance. (See Symmetric and Balanced Loading, above.)
1 To prevent microtiter plate breakage during centrifugation, place the flexible plastic pad,
ridged side up, into the flat, indented area of the blue base (see Figure 5.4).
2 Place the plate(s) on top of the pad, being careful not to spill the contents.
3 Slide the base, pad, and plate assembly into the carrier until the base clicks into place.
5-14 PN JR-IM-10AG
Using Swinging-Bucket Rotors
Rotor Preparation and Loading 5
Figure 5.4 The Micro Plus Microtiter Plate Carrier, Base, Pad, and Deep-Well Microtiter Plate
Deep-well Plate
Carrier
A
B
C
D
E
F
G
H
Base
Pad
NOTE When using stacked polypropylene plates, place a support pad beneath the bottom plate to prevent
breakage during centrifugation. Use the support pad beneath all polystyrene plates.
1 If using polystyrene or stacked polypropylene plates, place a support pad in the carrier with the
ridged-cross side down.
JS-5.3 Carrier
Support Pad
(369382)
Carrier
(368905)
PN JR-IM-10AG 5-15
Using Swinging-Bucket Rotors
Rotor Preparation and Loading
2 If using polystyrene or stacked polypropylene plates, place a support pad in the carrier with the
ridged-cross side down.
3 After centrifugation, grasp the carrier by the wire handles and lift it straight up out of the
bucket to unload it.
J6 Carriers
Carriers used with the JS-5.2 and JS-4.0 rotors are NOT interchangeable with those used with the
JS-4.2, JS-4.2A, JS 4.2SM, JS-3.0, and JS-2.9 rotors. If you have more than one type of carrier, check
the label on the side of the carrier to make sure that you are using the right one for your rotor.
Table 5.2 lists carriers used with J6 rotors.
1 To prevent microtiter plate breakage during centrifugation, place the rubber pad that comes
with each carrier on the bottom of the carrier.
Rubber
Pad
2 Place the plate(s) on top of the pad, being careful not to spill the contents.
5-16 PN JR-IM-10AG
Using Swinging-Bucket Rotors
Operation 5
Operation
Refer to the centrifuge instruction manual for detailed operating information. For low-temperature
runs, precool the rotor in the centrifuge or in a refrigerator before use—especially before short
runs—to ensure that the rotor reaches the set temperature. (To ensure that the rotor reaches the
required temperature during centrifugation, some temperature compensation may be required
because of the mass of these rotors. Refer to APPENDIX B or to the rotor manual for tables listing
temperature compensation units for various rotors.)
If you are using a microprocessor-controlled J2 or J6 series centrifuge, enter the rotor code (if the
JS-5.2 rotor is used for example, enter code 5.2).
Sample Recovery
CAUTION
If disassembly reveals evidence of leakage, you should assume that some fluid
escaped the container or rotor. Use appropriate decontamination procedures on
the centrifuge, rotor, and accessories.
1 Remove the rotor lid (if applicable). Remove the buckets or carriers from the rotor.
NOTE Except for the JS-24.38 and JS-24.15 rotors, you can leave the rotor body or yoke in the centrifuge
between runs unless spillage has occurred—in which case you should remove the buckets or carriers and
yoke and clean the centrifuge and rotor components immediately, according to the instructions in the
centrifuge and rotor instruction manuals. If the rotor is left in the centrifuge between runs, tighten the
tie-down device before each run. The JS-24.38 and JS-24.15 rotors must be removed from the centrifuge
to install or remove buckets.
1 Remove the rotor from the centrifuge by lifting it straight up and off the drive hub.
2 Set the rotor on the rotor stand and carefully remove the buckets.
PN JR-IM-10AG 5-17
Using Swinging-Bucket Rotors
Sample Recovery
3 Remove the bucket caps and use the appropriate removal tool (listed in the rotor manual) to
remove the spacers and tubes.
If floating spacers were used, remove them with the threaded end of the floating spacer
removal tool (338765).
While pressing
the rubber tip
against the
adapter wall, Extractor
pull the tool Tool
and adapter (354468)
up and out
of the cavity.
NOTE If the conical-shaped adapters that support konical tubes are difficult to remove after
centrifugation, an extractor tool (354468) is available to facilitate removal.
See CHAPTER 7 for instructions on the care of rotors, tubes or bottles, and accessories after a run.
5-18 PN JR-IM-10AG
CHAPTER 6
Using Vertical-Tube
and Rack-Type Rotors
Introduction
This section contains instructions for using vertical-tube or rack-type rotors in J series centrifuges. In
addition to these instructions, observe procedures and precautions provided in the applicable rotor and
centrifuge manuals.
Refer to CHAPTER 2 for tube selection information, and CHAPTER 3 for recommended filling and sealing
requirements for each tube type and for sample recovery procedures. Refer to CHAPTER 7 for information
on the care of rotors and accessories.
Description
Refer to Table 6.1 for general operating specifications for vertical-tube and rack-type rotors.
Table 6.1 General Specifications for Beckman Coulter J Series Vertical-Tube and Rack-Type Rotors
a. Maximum speeds are based on a solution density of 1.7 g/mL for the JV-20 rotor and 1.2 g/mL for the JR-3.2 rotor.
b. Critical speed range is the range of speeds over which the rotor shifts so as to rotate about its center of mass. Passing through or running
at the critical speed range is characterized by some vibration.
PN JR-IM-10AG 6-1
Using Vertical-Tube and Rack-Type Rotors
Description
Vertical-Tube Rotors
Vertical-tube rotors (see Figure 6.1) hold tubes parallel to the axis of rotation;
therefore, bands separate across the diameter of the tube rather than down the
length of the tube (see Figure 1.3). Vertical-tube rotors are useful for separating and
banding subcellular particles. These rotors have plugs that are screwed into the rotor
cavities over sealed Quick-Seal tubes. The plugs (with spacers, when required)
restrain the tubes in the cavities and provide support against the hydrostatic force
generated by centrifugation. Refer to CHAPTER 3 for information about filling and
sealing Quick-Seal tubes for use in vertical-tube rotors.
rmin
rav
rmax
Axis of Rotation
Rack-Type Rotors
The rack-type rotor (see Figure 6.2) holds a wide range of gamma-counter tubes in tube racks. Racks
are loaded into removable trays, which are then loaded into carriers at a resting angle. During
centrifugation, the carriers swing out to a completely horizontal position to provide uniform
pelleting of samples. The nearly vertical position of the racks during centrifugation permits
processing of up to 320 mL in one run.
rmin
rav
rmax
Axis of Rotation
6-2 PN JR-IM-10AG
Using Vertical-Tube and Rack-Type Rotors
Using a Vertical-Tube Rotor 6
NOTE Although rotor components and accessories made by other manufacturers may fit in the
Beckman Coulter rotor you are using, their safety in the rotor cannot be ascertained by Beckman Coulter.
Use of other manufacturers’ components or accessories in the Beckman Coulter rotor may void the rotor
warranty, and should be prohibited by your laboratory safety officer. Only the components and
accessories listed in the applicable rotor manual should be used.
1 Make sure that the rotor and plugs are clean and show no signs of corrosion or cracking.
2 Check the chemical compatibilities of all materials used. (Refer to APPENDIX A.)
3 Verify that the tubes, spacers, and floating spacers being used are in good condition and are
listed in the rotor manual.
Rotor Preparation
1 Be sure that the plug threads are lightly but evenly lubricated with Spinkote lubricant (306812).
2 Lubricate the rotor drive hole with silicone vacuum grease (335148).
PN JR-IM-10AG 6-3
Using Vertical-Tube and Rack-Type Rotors
Using a Vertical-Tube Rotor
3 Load the filled and sealed tubes symmetrically into the rotor.
Opposing tubes must be filled to the same level with liquid of the same density.
Refer to Rotor Balance in CHAPTER 1.
4 Insert spacers and floating spacers, as listed in the rotor manual, to completely fill rotor cavities
in use.
Spacer
Floating
Spacer
5 With the rotor in the rotor vise (332688), insert plugs over filled cavities only; do not insert plugs
in empty cavities.
Tighten the plugs using the plug wrench provided (340632).
Operation
Refer to the centrifuge instruction manual for detailed operating information. For low-temperature
runs, precool the rotor in the centrifuge or in a refrigerator before use—especially before short
runs—to ensure that the rotor reaches the set temperature. (To ensure that the rotor reaches the
required temperature during centrifugation, some temperature compensation may be required
because of the mass of these rotors. Refer to APPENDIX B or to the rotor manual for tables listing
temperature compensation units for various rotors.)
• If you are using an Avanti J series centrifuge, select the rotor number.
• If you are using a microprocessor-controlled J2 or J6 series centrifuge, enter the rotor code
(enter code 20 for the JV-20 rotor).
ROTOR
A/D
OPT
6-4 PN JR-IM-10AG
Using Vertical-Tube and Rack-Type Rotors
Using a Vertical-Tube Rotor 6
CAUTION
The centrifuge drive spindle can be bent or broken if the rotor is forced sideways
or dropped onto it. Install the rotor by centering it over the spindle and carefully
lowering it straight down.
1 Carefully lower the rotor straight down onto the drive spindle.
2 Rotate the rotor by hand until the drive pins seat on the drive spindle hub.
a. In older model centrifuges—be sure the pins in the rotor drive hole are located in the grooves
of the drive spindle hub.
Rotor
Drive Pins (2)
Hub
Grooves (4) Hub
Drive
Spindle
Assembly
b. In newer model centrifuges—be sure the pins in the rotor drive hole are not sitting on top of
the teeth on the drive spindle hub.
Teeth (2)
Hub
Drive
Spindle
Assembly
CAUTION
The pins located in the rotor hub must be seated correctly on the centrifuge drive
spindle. Running a rotor that is not seated properly may result in rotor failure.
3 Secure the rotor to the drive spindle hub with the rotor tie-down assembly.
PN JR-IM-10AG 6-5
Using Vertical-Tube and Rack-Type Rotors
Using a Vertical-Tube Rotor
CAUTION
If disassembly reveals evidence of leakage, you should assume that some fluid
escaped the container or rotor. Apply appropriate decontamination procedures to
the centrifuge and accessories.
2 If the rotor straight up and off the drive spindle. If the rotor sticks to the drive spindle, a rotor
removal tool may be used.
NOTE Lubrication of the centrifuge drive spindle hub with Spinkote should prevent the rotor from
sticking on all centrifuges except Avanti J series. Avanti J series centrifuges have acetal rings on the
spindle hubs to prevent sticking and do not require lubrication.
3 Place the rotor in the rotor vise and use the plug wrench to remove the rotor plugs.
Plug
Wrench
Rotor Vise
4 Remove spacers with the floating spacer removal tool (338765) and tubes with the tube removal
tool (361668).
6-6 PN JR-IM-10AG
Using Vertical-Tube and Rack-Type Rotors
Using a Rack-Type Rotor 6
Tray
Rack
1 Make sure that the rotor yoke and carriers are clean and show no signs of corrosion or cracking.
2 Check the chemical compatibilities of all materials used. (Refer to APPENDIX A.)
3 Verify that the tube racks, trays, adapters, and spacers being used are in good condition and are
listed in the rotor manual.
Rotor Preparation
1 Lubricate the rotor drive hole with silicone vacuum grease (335148).
2 Load racks into either two or four trays, then load tubes into the racks.
Do not over-fill tubes; leave enough space to avoid spills during carrier loading.
NOTE If all tubes to the same level with liquid of the same density. Racks and tubes must be horizontally
and vertically symmetrical during centrifugation.
PN JR-IM-10AG 6-7
Using Vertical-Tube and Rack-Type Rotors
Using a Rack-Type Rotor
Attachment
Knob
Carrier
Yoke
Operation
Refer to the centrifuge instruction manual for detailed operating information. For low-temperature
runs, precool the rotor in the centrifuge or in a refrigerator before use—especially before short
runs—to ensure that the rotor reaches the set temperature. (To ensure that the rotor reaches the
required temperature during centrifugation, some temperature compensation may be required
because of the mass of these rotors. Refer to APPENDIX B or to the rotor manual for tables listing
temperature compensation units for various rotors.)
If you are using a microprocessor-controlled J6 series centrifuge, enter the rotor code (enter
code 3.2 for the JR-3.2 rotor).
6-8 PN JR-IM-10AG
Using Vertical-Tube and Rack-Type Rotors
Using a Rack-Type Rotor 6
CAUTION
The centrifuge drive spindle can be bent or broken if the rotor is forced sideways
or dropped onto it. Install the rotor by centering it over the spindle and carefully
lowering it straight down.
1 Lift the rotor by the yoke and carefully lower it straight down onto the drive spindle.
2 Rotate the rotor by hand until the drive pins seat on the drive spindle hub.
• In older model centrifuges—be sure the pins in the rotor drive hole are located in the grooves
of the drive spindle hub.
Rotor
Drive Pins (2)
Hub
Grooves (4) Hub
Drive
Spindle
Assembly
• In newer model centrifuges—be sure the pins in the rotor drive hole are not sitting on top of
the teeth on the drive spindle hub.
Teeth (2)
Hub
Drive
Spindle
Assembly
CAUTION
The pins located in the rotor hub must be seated correctly on the centrifuge drive
spindle. Running a rotor that is not seated properly may result in rotor failure.
3 Turn the tie-down knob to the right (clockwise) until the rotor is secure.
PN JR-IM-10AG 6-9
Using Vertical-Tube and Rack-Type Rotors
Using a Rack-Type Rotor
CAUTION
If disassembly reveals evidence of leakage, you should assume that some fluid
escaped the container or rotor. Apply appropriate decontamination procedures to
the centrifuge, rotor, and accessories.
1 Turn the tie-down knob to the left (counterclockwise) to release the rotor from the drive
spindle.
NOTE Lubrication of the centrifuge drive spindle hub with Spinkote should prevent the rotor from
sticking.
3 To remove a tray from a carrier, lift the end of the tray just enough to clear the carrier.
Slide the tray up so that it passes under the hinge pins.
6-10 PN JR-IM-10AG
CHAPTER 7
Introduction
This section provides information on the care of rotors and accessories. Included is a list of some common
operating problems with suggestions for their solutions. Rotors and accessories should be kept in optimal
condition, thus minimizing the chances of rotor or labware failure. In addition to these instructions,
observe procedures and precautions provided in individual rotor manuals. Chemical Resistances (IN-175)
provides the chemical compatibilities of rotor and accessory materials to various acids, bases, salts, and
solvents.
Rotor Care
Rotor care involves not only careful operating procedures but also careful attention to:
• Regular cleaning, decontamination, and/or sterilization as required,
• Frequent inspection,
• Corrosion prevention, and
• Regular and proper lubrication.
Do not use sharp tools on a rotor, as the surface can get scratched. Corrosion begins in scratches and
may open fissures in the rotor with continued use. The corrosion process accelerates with speed-
induced stresses. The potential for damage from corrosion is greatest in aluminum rotors and
components.
PN JR-IM-10AG 7-1
Care and Maintenance
Rotor Care
Cleaning
Wash rotors and rotor components immediately if salts or other corrosive materials are used or if spillage
has occurred. DO NOT allow corrosive materials to dry on the rotor.
NOTE Do not wash rotor components or accessories in a dishwasher. Do not soak in detergent solution for
long periods, such as overnight.
With normal usage, wash rotors frequently to prevent corrosion that can begin in scratches.
Remove buckets from yokes before cleaning swinging-bucket rotors.
CAUTION
Do not immerse or spray a swinging bucket rotor yoke (or body) with water
because liquid can become trapped in the hinge pin area and lead to corrosion.
1 Use plastic or wooden tools to remove O-rings or gaskets for cleaning—do not use metal tools
that could scratch anodized surfaces.
a. Use a mild detergent such as Solution 555 (339555) and a soft brush to wash rotors and rotor
components and accessories.
• Dilute the detergent with water (10 parts water to 1 part detergent).
• (Most laboratory detergents are too harsh for aluminum rotors and components.)
• The Rotor Cleaning Kit (339558) contains two quarts of Solution 555 and brushes that
will not scratch rotor surfaces.
Rotor Cleaning
Kit (339558)
7-2 PN JR-IM-10AG
Care and Maintenance
Rotor Care 7
6 Approximately once a week (or every 250 runs), clean the pins and bucket pin sockets of
swinging bucket rotors to prevent buildup of residues.
After cleaning, lubricate pin sockets as described under Lubrication, below.
CAUTION
Do not use acetone, MEK (methylethylketone), chloroform, cyclohexane, or
organic solvents on carbon-fiber canisters at any time. These substances will
damage the epoxy resin surface material.
Decontamination
Rotors contaminated with radioactive or pathogenic materials must be decontaminated, following
appropriate laboratory safety guidelines and/or other regulations.
NOTE. Strong bases and/or high-pH solutions can damage aluminum rotors and components.
• If a rotor (and/or accessories) becomes contaminated with radioactive material, it should be
decontaminated using a solution that will not damage the anodized surfaces. Beckman Coulter
has tested a number of solutions and found two that do not harm anodized aluminum: RadCon
Surface Spray or IsoClean Solution (for soaking), *and Radiacwash.†
NOTE. IsoClean can cause fading of colored anodized surfaces. Use it only when necessary, and do not soak
rotor components longer than the minimum time specified in the IsoClean usage instructions. Then
remove it promptly from surfaces.
While Beckman Coulter has tested these methods and found that they do not damage
components, no guarantee of decontamination is expressed or implied. Consult your laboratory
safety officer regarding the proper decontamination methods to use.
• If the rotor or other components are contaminated with toxic or pathogenic materials, follow
appropriate decontamination procedures as outlined by appropriate laboratory safety
guidelines and/or other regulations. Consult Chemical Resistances (IN-175) to select an agent
that will not damage the rotor.
* In U.S., contact Nuclear Associates (New York); in Eastern Europe and Commonwealth States, contact Victoreen GmbH
(Munich); in South Pacific, contact Gammasonics Pty. Ltd. (Australia); in Japan, contact Toyo Medic Co. Ltd. (Tokyo).
† In U.S., contact Biodex Medical Systems (Shirley, New York); internationally, contact the U.S. office to find the dealer
closest to you.
PN JR-IM-10AG 7-3
Care and Maintenance
Rotor Care
• Rotors and most rotor components (except those made of modified polyphenylene oxide) can
be autoclaved at 121°C for up to an hour. Remove the lid and place the rotor (and/or buckets)
in the autoclave upside-down. (O-rings and gaskets can be left in place on the rotor.)
• Ethanol (70%) may be used on all rotor components, including those made of plastic. Bleach
(sodium hypochlorite) may be used, but may cause discoloration of anodized surfaces. Use the
minimum immersion time for each solution, per laboratory standards.
CAUTION
Ethanol is a flammability hazard. Do not use it in or near operating centrifuges.
Inspection
Frequent and thorough inspection is crucial to maintaining a rotor in good operating condition.
• Periodically (at least monthly, depending on use) inspect the rotor, especially inside cavities
and buckets, for rough spots, cracks, pitting, white powder deposits on aluminum rotors
(frequently aluminum oxide), or heavy discoloration. If any of these signs are evident, do not
run the rotor. Contact your Beckman Coulter representative for information about the Field
Rotor Inspection Program and the Rotor Repair Program.
• Regularly check the condition of O-rings or gaskets and replace any that are worn or damaged.
• Regularly check that all sealing surfaces are smooth and undamaged to ensure proper sealing.
7-4 PN JR-IM-10AG
Care and Maintenance
Rotor Care 7
Sealing
Surface
Before each use, inspect carbon-fiber canisters for cracks
Aluminum- S/N XXXXX
X / Exp. Date 04/99
Made in USA
where carbon-fiber threads are visible. If any cracking or
Composite
other damage is visible on the outside wall or near the
JLA-10.500 10,000 RPM
Interface
aluminum-composite interface area, do not use the
Outside
Wall
Sleeve
Washer
canister. Contact your Beckman Coulter representative.
Retire the canister on the expiration date.
Lubrication
Regular and thorough lubrication can extend the useful life of rotor components.
O-Rings
Many rotors use O-rings or gaskets as seals to maintain atmospheric pressure in the rotor during
a run. These O-rings and the rotor surfaces they bear against must be kept clean and evenly
lubricated. After removing and cleaning rotor or bucket O-rings or gaskets, lightly but uniformly
coat them with silicone vacuum grease and reposition them in the rotor.
NOTE Do not apply lubricant with a cotton-tipped swab. These swabs can leave lint on the O-ring or gasket
that can interfere with the seal.
PN JR-IM-10AG 7-5
Care and Maintenance
Tube, Bottle, and Accessory Care
• to prevent premature rotor failures by detecting conditions such as stress, corrosion, metal
fatigue, damage, or wear in the anodized coatings; and
• to instruct laboratory personnel in the proper care of rotors.
Beckman Coulter has trained a group of experienced service engineers in the techniques
of nondestructive evaluation. For more information about the program, contact your
Beckman Coulter representative.*
Proper care of tubes and bottles involves observing temperature, fill volume, and run speed
limitations as well as careful cleaning and sterilization procedures.
Cleaning
Do not wash tubes and bottles in a commercial dishwasher —detergents and temperatures are too harsh.
• Wash tubes, bottles, adapters, and blood bag cups by hand, using a
mild detergent, such as Solution 555 (339555) and a soft brush.
• Dilute the detergent with water (10 parts water to 1 part detergent).
• Disassemble multitube adapters for cleaning. After washing with
Solution 555 and a soft brush, rinse them with water, then dry and
reassemble.
• Polycarbonate bottles and tubes are vulnerable to attack by alkaline
solutions and detergents, so use a detergent with pH less than 9, such
as Solution 555. Do not use a brush with exposed metal; scratches in
polycarbonate will cause early failure.
• Alcohol and acetone react unsatisfactorily with many tube and accessory materials. If a solvent
must be used to rinse, dry, or decontaminate these materials, consult APPENDIX A to select an
appropriate solvent.
• Do not dry tubes, bottles, or accessories in an oven. Labware should be air-dried.
• Quick-Seal, Ultra-Clear, and thinwall polypropylene tubes are intended for one-time use and
should be discarded after use.
* Call 1-800-742-2345 (U.S.A. or Canada), outside the U.S., contact your local Beckman Coulter office.
7-6 PN JR-IM-10AG
Care and Maintenance
Tube, Bottle, and Accessory Care 7
Decontamination
CAUTION
Do not autoclave tubes or bottles with caps on. Pressure in a sealed container can
cause an explosion. Pressures within the autoclave can cause partially sealed
containers to collapse when the autoclave vents.
PN JR-IM-10AG 7-7
Care and Maintenance
Tube, Bottle, and Accessory Care
Sodium
Hypo- Hydrogen Glutaral- Phenolic
Tube/Bottle Autoclaveb UV Ethylene Formal- Ethanol chlorite Peroxide dehyde Deriva-
Material (121°C) Irradiation Oxide dehyde (70%) c (10%) (10%) (2%) tives
polypropylene yes no yes yes yes yes yes yes no
Ultra-Clear no no yes yesd yes yes yes yes no
polycarbonate yese no yes yesd no yesf yes yes no
polypropylene yes no yes yes yes yesg yesh yes no
polyethylene no no yes yes yeis yes yes yes yes
cellulose no no no no no yes yes yes no
propionate
stainless steel yes yes yes yes yesj no yes yes no
a. This information is provided as a guide to the use of sterilization and disinfection techniques for tube materials. Cold sterilization results shown are
for short-duration (10-minute) soak periods; reactions may differ with extended contact. Refer to Appendix A of this manual for information about
specific solutions.
b. To avoid deformation, autoclave tubes or bottles open-end down in a tube rack at 15 psig for no more than 20 minutes (allow to cool before removing
from tube rack). DO NOT autoclave capped or sealed tubes or bottles.
c. Flammable; do not use in or near operating centrifuges.
d. Do not use if there is methanol in the formula.
e. Tube life will be reduced by autoclaving.
f. Discoloration may occur.
g. Can be used if diluted.
h. Below 26°C only.
i. Below 21°C only.
j. Marginal.
JS-5.0 labware cups, cup covers, cup gaskets (369261 and 369257), and partitions can be autoclaved
at 121°C for up to 20 minutes. Remove the plug and air-vent filter from each cup cover before
autoclaving, and remove the gasket from the cup. To remove an air-vent filter, gently push it out
from underneath the cover with a pencil or other non-metal tool that will not scratch the cover
material. After autoclaving, insert a new air-vent filter into each cup cover. Thoroughly dry the
gasket sealing surfaces before replacing the gasket.
CAUTION
Autoclaving will reduce the useful life of the labware cups, cup covers, cup
gaskets, and partitions. After each autoclave cycle, examine these components
for damage and DO NOT USE damaged components.
HarvestLine system liners can be gamma irradiated to a maximum dose of 40.0 kGy. Gamma
irradiation causes the liners to become yellow, but does not affect their performance. Do not steam
or dry autoclave the liners or they will be damaged. The liners are designed for single use only.
A cold sterilization method, such as immersion in 10% hydrogen peroxide for 30 minutes, may be
used on Ultra-Clear tubes. Refer to Table 7.1 to select cold sterilization materials that will not
damage tubes and accessories.
7-8 PN JR-IM-10AG
Care and Maintenance
Tube, Bottle, and Accessory Care 7
While Beckman Coulter has tested these methods and found that they do not damage the
components, no guarantee of sterility or disinfection is expressed or implied. When sterilization or
disinfection is a concern, consult your laboratory safety officer regarding proper methods to use.
Inspection
Inspect containers and accessories before use.
• Inspect tubes and bottles for cracks or any major deformities before using them.
• Do not use a tube that has become yellowed or brittle with age or excess exposure to ultraviolet
light.
• Crazing—the appearance of fine cracks on tubes and bottles—is the result of stress relaxation.
If a crack approaches the outer wall of the tube or bottle, discard it.
Crazing
Cracking
NOTE Replace the air-vent filter in each JS-5.0 cup cover after every 250 cycles, or after every
autoclave cycle.
Air-Vent Filter
PN JR-IM-10AG 7-9
Care and Maintenance
Returning a Rotor or Accessory to the Factory
Before returning a rotor or accessory for any reason, prior permission must be obtained from
Beckman Coulter, Inc. The return authorization form may be obtained from your local
Beckman Coulter sales office. The form, entitled Returned Material Authorization (RMA) for
United States returns or Returned Goods Authorization (RGA) for international returns, should
contain the following information:
To protect our personnel, it is the customer’s responsibility to ensure that the parts are free from
pathogens, chemical hazards, and/or radioactivity. Sterilization and decontamination MUST be
done before returning the parts. Smaller items (such as tubes, bottles, and so on) should be enclosed
in a sealed plastic bag.
All parts must be accompanied by a note, plainly visible on the outside of the box or bag, stating that they
are safe to handle and that they are not contaminated with pathogens, chemical hazards, or radioactivity.
Failure to attach this notification will result in return or disposal of the items without review of the
reported problem.
Use the address label printed on the RMA/RGA form when mailing the rotor and/or accessories.
Customers located outside the United States should contact their local Beckman Coulter office.
Diagnostic Hints
Some of the more common operating problems experienced in centrifugation are listed in Table 7.2 with
suggestions for their solutions. Contact your Beckman Coulter Field Service representative if a problem
cannot be corrected.
NOTE Use only the labware listed in the applicable rotor manual.
7-10 PN JR-IM-10AG
Care and Maintenance
Diagnostic Hints 7
Rotor
Severe vibration • Rotor imbalance. To balance the rotor load, fill all opposing tubes to
the same level with liquid of the same density. Weight of opposing
tubes must be distributed equally. Place tubes in a fixed-angle or
vertical-tube rotor symmetrically, as illustrated in CHAPTER 1
(Figure 1.6). Detailed information about balancing swinging-bucket
rotors is contained in CHAPTER 5.
Rotor lid, canister cover, or bucket cover is • Vacuum built up inside the container during centrifugation. Lift the
difficult to remove after centrifugation vent plug on bucket or canister covers with vents to relieve the
vacuum.
• Threads contaminated with dirt, dried lubricant, or metal particles, or
threads insufficiently lubricated cause rotor components to stick.
Do not use excessive force to loosen components. Contact your
Beckman Coulter representative. Routinely clean metal threads with
concentrated Solution 555 that has been diluted to 10 parts water to
1 part detergent, then lubricate them with Spinkote.
Anodizing coming off where bucket or Not an operational problem (some buckets are not anodized inside the
carrier contacts rotor pins on swinging- pin pockets to facilitate swinging).
bucket rotor
Adapter
Adapters stick in buckets after Apply a thin film of powder, such as talcum powder, to the tube adapter
centrifugation rubber bases after cleaning or as required to prevent sticking.
Tube
Tube leakage
Tubes with cap assemblies • Caps not properly secured. Caps must be properly seated on tubes
and then fully tightened.
• Cap components not dry before assembly. Thoroughly dry all
components before assembling.
PN JR-IM-10AG 7-11
Care and Maintenance
Diagnostic Hints
Tubes with snap-on caps Tube too full; the meniscus must be kept lower to prevent leakage.
Uncapped tubes Tube volume exceeds maximum uncapped volume. Refer to the rotor
manual for tube volumes and speed reductions.
Quick-Seal tubes Improperly sealed. After heat-sealing, squeeze the tube gently (if the
tube contents may be disturbed) to test the seal for leaks. If the tube
leaks, reseal it.
Tube cracking • Tubes may crack or become brittle if they are used below their lower
temperature limit. Before using tubes at other than stated
temperature limits, evaluate them under centrifugation conditions. If
sample is frozen in tubes, make sure that they are thawed to at least
2°C before centrifugation.
• Tubes may become brittle with age and use. Dispose of brittle or
cracked tubes.
Tube collapse • Thinwall tube volume too low to provide tube wall support. Meniscus
should be 2 to 3 mm below the tube top. Refer to the rotor manual for
tube volumes.
• Moisture between the tube and the cavity or bucket can cause the
tube to float and collapse. Ensure that tubes and tube cavities or
buckets are dry before inserting the tubes.
• Reagent used that attacks the tube material. Refer to Chemical
Resistances (IN-175) for chemical compatibilities of tube material and
chemicals.
• Tubes run above their rated speed. Refer to the applicable rotor
manual for maximum speeds.
Bottle
Bottle leakage (bottles with cap assemblies) • Moisture or lubrication on cap or sealing surface. Ensure that the
O-ring, plug, and bottle lip are dry and free of lubrication before use.
• O-ring or gasket damaged or defective. Replace the O-ring or gasket.
• Cap not tightened sufficiently. Tighten cap securely.
• Sealing surface of the bottle is not smooth. Replace bottle.
• Threaded caps without inserts or O-rings—Tube too full; these are not
as liquid-tight as cap assemblies; therefore, the meniscus must be
kept lower to prevent leakage.
7-12 PN JR-IM-10AG
Care and Maintenance
Diagnostic Hints 7
Bottle leakage (uncapped bottles) Bottle too full; the meniscus must be kept lower to prevent leakage. Refer
to the rotor manual for fill volumes and speed reductions.
Bottle damage • Fill volume too low to provide tube wall support. Refer to the rotor
manual for fill volumes and speed reduction.
• Moisture between the bottle and the cavity or bucket can cause the
bottle to float and collapse. Ensure that bottles and cavities or buckets
are dry before inserting them.
• Reagent used that attacks the bottle material. Refer to Chemical
Resistances (IN-175) for chemical compatibilities of bottle material
and chemicals.
• Bottles may crack or become brittle if they are used below their lower
temperature limit. Before using bottles at other than stated
temperature limits, evaluate them under centrifugation conditions. If
sample is frozen in bottles, make sure that they are thawed to at least
2°C before centrifugation.
• Bottles may become brittle with age and use. Dispose of brittle or
cracked bottles.
• Improper cleaning, decontamination, or sterilization procedures used.
Refer to Table 7.1 for acceptable procedures and materials.
a. Critical speed range is the range of speeds over which the rotor shifts so as to rotate about its center of mass. Passing through the critical speed
range is characterized by some vibration.
PN JR-IM-10AG 7-13
Care and Maintenance
Diagnostic Hints
7-14 PN JR-IM-10AG
APPENDIX A
PN JR-IM-10AG A-1
Chemical Resistances for Beckman Coulter Centrifugation Products
List of Chemical Resistances
A-2 PN JR-IM-10AG
APPENDIX B
Introduction
This Appendix contains tables listing temperature compensation units for various rotors used in
Beckman Coulter discontinued J2 and J6 series centrifuges.
Temperature Compensation
To ensure that the rotor reaches the required temperature during centrifugation, some
temperature compensation may be required because of the mass of these rotors. The following
tables list temperature compensation units for various rotors.
Refer to CHAPTER 1 or the applicable rotor manual for procedures to set the temperature
compensation for the model of J centrifuge being used.
NOTE When using an Avanti J series centrifuge, enter the run temperature according to the instructions in
your centrifuge instruction manual. No additional input is required.
18,000 –8 –7 –5 –6 –6 –5 +5
JA-25.15 15,000 –6 –5 –4 –4 –3 –4 +6
10,000 –3 –2 –1 –2 –1 –1 +9
18,000 N –8 –8 –7 –6 –5 –3
JA-21 15,000 –6 –6 –5 –4 –4 –3 N
10,000 –4 –2 –1 –1 –1 –1 N
18,000 N N –7 –7 –7 –6 –4
JA-20.1 15,000 –7 –5 –5 –4 –4 –3 –2
10,000 –4 –2 –1 –1 –1 –1 N
PN JR-IM-10AG B-1
Temperature Compensation Tables
Temperature Compensation
16,000 N N –8 –5 –4 –3 0
15,000 N N –2 –2 –2 –2 0
JA-18.1c
10,000 –4 –3 –3 –3 –3 –2 N
8,000 –1 –2 –4 –3 –2 –1 N
18,000 N N N N N –8 –4
17,000 N N N – 10 –9 –8 –3
16,000 N N – 10 –9 –8 –7 –2
JA-18
12,000 N –6 –6 –5 –4 –3 –1
8,000 –3 –2 –2 –1 –1 0 N
5,000 –1 –1 –1 –1 0 +1 N
17,000 N N –6 –6 –6 –5 –3
15,000 N –5 –4 –4 –4 –3 –2
JA-17
12,000 –4 –4 –3 –3 –3 –2 N
8,000 –2 –2 –2 –2 –2 –1 N
14,000 N N 7 –7 –7 –5 –4
JLA-16.250 10,000 N –4 –3 –3 –2 –1 N
5,000 N 0 0 –3 –3 –3 N
14,000 N N –5 –5 –5 –3 –2
12,000 N –3 –3 –3 –3 –2 –1
JA-14
10,000 –4 –2 –2 –2 –1 –1 N
5,000 –1 0 0 0 0 0 N
12,000 N –3 –2 –2 –2 –2 –1
JA-12 10,000 –2 –2 –1 –1 –1 –1 –1
5,000 –1 –1 0 0 0 0 0
10,000 N –3 –2 –1 0 +1 +2
JA-10 8,000 –3 –2 0 0 0 +1 +2
5,000 –2 0 0 0 0 +1 +2
10,000 N – 6d –3 –3 –3 –1 0
JLA-10.500 8,000 – 3† –3 –3 –2 –2 –1 0
5,000 – 3† –2 –2 –1 0 0 +2
13,000 N –6 –9 –9 –9 –6 –3
11,000 N –5 –5 –5 –5 –4 –1
JS-13.1
8,000 –5 –3 –2 –2 –1 –1 N
5,000 –4 –2 –1 –1 –1 0 N
7,500 0 +2 +3 +3 +4 +4 +4
JS-7.5 5,000 0 +1 +2 +2 +2 +3 +3
2,000 0 0 0 +1 +1 +2 N
B-2 PN JR-IM-10AG
Temperature Compensation Tables
Temperature Compensation B
Table B.2 Temperature Compensation Settings for the J2-21, J2-21B, J2-21C, and J2-HS Centrifugesa
PN JR-IM-10AG B-3
Temperature Compensation Tables
Temperature Compensation
Table B.2 Temperature Compensation Settings for the J2-21, J2-21B, J2-21C, and J2-HS Centrifugesa (Continued)
B-4 PN JR-IM-10AG
Temperature Compensation Tables
Temperature Compensation B
Table B.3 Temperature Compensation Settings for the J2-MI, J2-21M, J2-MC, and J2-21M/E Centrifuges
20,000 Na N –7 –5 –6 –5 –9
18,000 N –5 –5 –5 –5 –4 –7
JA-25.50
15,000 0 –2 –3 –3 –2 –2 –6
10,000 0 0 –1 0 –1 0 –3
21,000 N N –8 –8 –7 –6 –9
18,000 N –8 –8 –8 –4 –5 –5
JA-25.15
15,000 –6 –6 –5 –4 –4 –3 –2
10,000 –3 –2 –1 –1 –1 –1 –2
21,000 N N – 10 – 10 – 10 –8 –6
18,000 N –8 –8 –7 –6 –5 –3
JA-21
15,000 –6 –6 –5 –4 –4 –3 N
10,000 –3 –2 –1 –1 –1 0 N
20,000 N N – 10 –9 –9 –9 –7
18,000 N N –7 –7 –7 –6 –4
JA-20.1
15,000 –7 –5 –4 –4 –4 –3 N
10,000 –2 –1 –1 0 0 0 N
20,000 N N N 11 10 9 –6
18,000 N N 8 7.5 7 6 –3
JA-20
15,000 –5 –4 4 4 3 3 –1
10,000 –2 –2 2 1 1 0 0
18,000 N N N N –8 –5 0
17,000 N N N N –6 –5 0
JA-18.1 15,000 N N –9 –5 –4 –3 0
10,000 –7 –7 –3 –3 –2 0 +2
8,000 –6 –5 –3 –3 –1 0 +2
18,000 N N N N N – 10 –7
17,000 N N N – 10 –9 –8 –6
16,000 N N – 10 – 10 –9 –7 –5
JA-18
12,000 N –6 –6 –6 –5 –4 –3
8,000 –5 –4 –4 –3 –2 –1 0
5,000 –4 –3 –2 –2 –1 0 0
17,000 N N –7 –7 –7 –4 –2
15,000 N –5 –4 –4 –4 –3 –1
JA-17
12,000 –4 –4 –3 –3 –3 –1 N
8,000 –1 –1 0 0 0 0 N
14,000 N N –7 –5 –4 –3 –3
JLA-16.250 10,000 N –4 –3 –3 –2 –1 N
5,000 N 0 0 0 0 0 N
14,000 N N –7 –7 –7 –5 –4
12,000 N –6 –5 –5 –4 –3 –2
JA-14
10,000 –4 –4 –3 –3 –2 –1 N
5,000 –1 0 0 0 0 0 N
PN JR-IM-10AG B-5
Temperature Compensation Tables
Temperature Compensation
Table B.3 Temperature Compensation Settings for the J2-MI, J2-21M, J2-MC, and J2-21M/E Centrifuges (Continued)
12,000 N –6 –6 –7 –7 –6 –9
JA-12 10,000 –3 –4 –5 –5 –5 –5 –9
5,000 –1 –3 –3 –4 –4 –5 –9
10,000 N –3 –2 –1 0 +1 +2
JA-10 8,000 –3 –2 –1 –1 0 +1 +2
5,000 –2 0 0 0 0 +1 +2
10,000 N – 5b –2 –1 0 +2 +1
JLA-10.500 8,000 – 1b – 1b 0 +2 +2 +3 +1
5,000 – 0* –1 +2 +1 +4 +3 +1
13,000 N – 10 – 10 –10 – 10 – 10 –9
11,000 N –8 –8 –8 –7 –6 –5
JS-13.1
8,000 –6 –5 –4 –4 –3 –3 N
5,000 –4 –3 –1 –1 –1 –1 N
7,500 –4 –3 +1 +1 +1 +2 +3
JS-7.5 5,000 –2 0 +3 +3 +3 +3 +4
2,000 0 +2 +4 +4 +4 +4 +5
20,000 N N N N – max – max – max
18,000 N N N – max – 10 – 10 –9
JV-20
15,000 N N –7 –7 –6 –5 –4
10,000 N N –2 –2 –1 0 0
a. N: indicates that the rotor cannot achieve the required temperature at this speed.
b. Above 30°C ambient temperature, this temperature may not be achievable at this speed.
5200 –3 –1 4 6 10 17 28
4000 –1 1 5 7 12 19 29
JS-5.2 3000 0 2 7 9 14 20 30
2000 and 2 4 8 10 15 20 30
below
JS-4,2, 4200 –3 0 5 7 13 19 30
JS-4.2SM, 3000 0 2 7 9 14 20 30
JS-4.2A, 2000 and 2 4 8 10 15 20 30
JS-4.2SMA below
B-6 PN JR-IM-10AG
APPENDIX C
Gradient Materials
Introduction
This Appendix contains reference information on commonly used gradient materials. General instructions
for filling and sealing tubes, including gradient preparation, are contained in CHAPTER 3.
Gradient material selection depends on a number of factors, including the type of separation to be
performed. Sucrose is used for rate zonal and isopycnic separations, and cesium chloride is often
used for isopycnic separations. The basic requirement is that the gradient permit the type of
separation. Additional considerations in selecting a gradient material include the following.
• Its density range should be sufficient to permit separation of the particles of interest by the
chosen density gradient technique, without overstressing the rotor.
• It should not affect the biological activity of the sample.
• It should be neither hyperosmotic or hypoosmotic when the sample is composed of sensitive
organelles.
• It should not interfere with the assay technique.
• It should be removable from the purified product.
• It should not absorb in the ultraviolet or visible range.
• It should be inexpensive and readily available; more expensive materials should be recoverable
for reuse.
• It should be sterilizable.
• It should not be corrosive to the rotor.
• It should not be flammable or toxic to the extent that its aerosols could be hazardous.
The following charts are provided as a reference for information on commonly used gradient
materials.
PN JR-IM-10AG C-1
Gradient Materials
Introduction
Maximum
Density
Materials Solvent at 20°C
C-2 PN JR-IM-10AG
Gradient Materials
Introduction C
Table C.2 Density, Refractive Index, and Concentration Data—Cesium Chloride at 25°C, Molecular Weight = 168.37a
PN JR-IM-10AG C-3
Gradient Materials
Introduction
Table C.3 Density, Refractive Index, and Concentration Data—Sucrose at 20°C, Molecular Weight = 342.3a
C-4 PN JR-IM-10AG
Gradient Materials
Introduction C
Table C.4 Density Conversion for Cesium and Rubidum Salts at 20°C
% w/w CsCl CsBr Csl Cs2SO4 CsNO3 RbCl RbBr Rbl Rb2SO4 RbNO3
1 1.00593 1.00612 1.00608 1.0061 1.00566 1.00561 1.00593 1.00591 1.0066 1.0053
2 1.01374 1.01412 1.01402 1.0144 1.01319 1.01307 1.01372 1.01370 1.0150 1.0125
4 1.02969 1.03048 1.03029 1.0316 1.02859 1.02825 1.02965 1.02963 1.0322 1.0272
6 1.04609 1.04734 1.04707 1.0494 1.04443 1.04379 1.04604 1.04604 1.0499 1.0422
8 1.06297 1.06472 1.06438 1.0676 1.06072 1.05917 1.06291 1.06296 1.0680 1.0575
10 1.08036 1.08265 1.08225 1.0870 1.07745 1.07604 1.08028 1.08041 1.0864 1.0731
12 1.09828 1.10116 1.10071 1.1071 1.09463 1.09281 1.09817 1.09842 1.1052 1.0892
14 1.11676 1.12029 1.11979 1.1275 1.11227 1.11004 1.11661 1.11701 1.1246 1.1057
16 1.13582 1.14007 1.13953 1.1484 1.12775 1.13563 1.13621 1.1446 1.1227
18 1.15549 1.16053 1.15996 1.1696 1.14596 1.15526 1.15605 1.1652 1.1401
PN JR-IM-10AG C-5
Gradient Materials
Introduction
C-6 PN JR-IM-10AG
APPENDIX D
Introduction
This Appendix provides a basic overview of blood separation procedures using Beckman Coulter J series
centrifuges.
Blood is composed of plasma, red blood cells (RBC), white blood cells (WBC), and platelets.
Approximately 40 to 45 percent of this volume is made up of red blood cells; most of the remainder
is plasma, a watery substance that contains vital substances, including hormones and proteins.
Most whole blood collected undergoes fractionation, or separation into components, in order to use
collected blood most efficiently. Termed “component therapy,” multiple use of different parts of
the blood helps conserve this scarce resource and allows patients to receive only the components
they need. As shelf life and storage requirements vary, conditions can be optimized by separating
components.
In addition to collection of whole blood for separation into components, several techniques involve
collection of whole blood, separation, collection of a fraction for infusion, and return of the
remainder to the donor.
• In plasmapheresis a unit of blood is taken to obtain plasma, separated, and the red cells are
immediately reinfused to the donor. Most plasmapheresis is performed for “source plasma,”
which is not intended for intravenous transfusion, but separated by large-scale fractionation
into clotting factors (especially factor VIII), albumin, and specific immunoglobulins.
• During plateletpheresis, whole blood is collected, platelets separated via centrifugations, and
platelet-depleted red blood cells returned to the donor. The plasma may be returned to the
donor or collected for fractionation into clotting factors and albumin.
• Leukapheresis is the separation of leukocytes, or white blood cells, from whole blood. The
leukocyte-depleted and platelet-depleted red blood cells are continuously or intermittently
returned to the donor.
PN JR-IM-10AG D-1
Blood Component Separation
Components and Typical Usage
• Fresh plasma—used for the treatment of deficiencies of clotting factors V, XI, and XIII.
• Factor VII Concentrate—separated from platelet-rich plasma, useful for treating clotting
deficiencies other than those mentioned above.
• Cryoprecipitated Antihemophilic Factor (factor VIII)—a protein concentrate separated from
cell-free plasma (frozen, then thawed at 4°C), useful for treating hemophilia.
• Platelet Concentrates—separated from plasma, platelet concentrates are used to treat
decreased platelet counts or failing platelet functions. Platelets must be separated by
centrifugation within 6 hours of collection.
Single-Donor Plasma
Single-donor plasma can be separated from whole blood up to a few days after the expiration date,
since no attempt is made to maintain the activity of the labile clotting factors. This plasma, which
may be frozen and kept for up to 5 years, is used for expansion of blood volume (treatment of
hypovolemic shock, caused by a dangerous reduction in blood volume).
D-2 PN JR-IM-10AG
Blood Component Separation
Shelf Life D
Shelf Life
Table D.1 lists approximate storage times for the separated components.
Approximate
Component Storage Life
Plasma—frozen 1 year
a. ADSOL = dextrose-sodium chloride-mannitol-adenine.
b. SAG-M = saline-adenine-glucose-minitol.
c. CPDA-1 = citrate-phosphate-dextrose-citrate-citric acid-adenine.
Freezing
A cryoprotective agent such as glycerol can be added to extend the life of frozen red blood cells.
These cells can then be stored for up to 3 years at –80°C. Prior to use the cells are thawed and the
glycerol is removed by washing.
If the plasma will not be separated within 15 hours of collection, it must be frozen within 6 hours of
collection. Freezing must be carried out in a flash freezer with complete freezing accomplished
within 1 or 2 hours of the time it is placed in the freezer.
Centrifugation is the primary method for processing blood because it offers the required high
throughput, reproducibility, and versatility. Most blood components can be separated in one or
two runs. Generally, two types of centrifugation runs are performed (see Figure D.1). Soft spin runs,
short centrifugation runs (3 to 5 minutes) at low g-forces (2000 to 3000 × g) at ambient temperature,
are used to keep small cells or platelets in suspension while the larger cells sediment. This type of
run is used to obtain platelet-rich plasma and red blood cell concentrate from whole blood.
Hard spin runs are longer (5 to 7 minutes), at higher g-forces (4000 to 5000 × g), at ambient
temperature or at 4°C, and are used to separate fresh plasma from cellular components. Soft spin
and hard spin techniques are often combined.
PN JR-IM-10AG D-3
Blood Component Separation
Separation of Blood Components by Centrifugation
Donor blood is collected in plastic bags with one or more satellite bags
(double, triple, or quad packs) containing anticoagulant and preservative.
After each centrifugation run, the sedimented fraction is squeezed into its
respective satellite bag. Common anticoagulants and preservatives
include citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose-
citrate-citric acid-adenine (CPDA-1), saline-adenine-glucose-minitol
(SAG-M), and dextrose-sodium chloride-mannitol-adenine (ADSOL).
hard spin
Platelet-rich Plasma
Plasma (Platelet-depleted)
soft spin
Red Blood Cells
Platelet
Concentrate
Whole Blood
slow-thaw,hard
eze,
fre
t
Plasma
(Cell-Free) (Cryoprecipitate
Removed)
Packed Cells
Cryoprecipitated
Antihemophilic
Factor (Factor VIII)
Beckman Coulter has centrifuges, rotors, and accessories designed to fit the special needs of blood
component processing. Several rotors are available to accommodate single, double, triple, and quad
blood bags. Blood bag cups rest in the rotor bucket and simplify processing, since they eliminate the
need to remove buckets after each run. They also minimize clean-up downtime if a bag breaks—
simply remove the cup and resume the run. Refer to the applicable rotor manual for blood bag cups
used with each rotor. Table D.2 lists blood bank methods that can be used for separating components
in a variety of J6 series centrifuges.
D-4 PN JR-IM-10AG
Blood Component Separation
Separation of Blood Components by Centrifugation D
PN JR-IM-10AG D-5
Blood Component Separation
Tips for Optimum Centrifugation Runs
Centrifugation generates high speeds, causing rotor heads and buckets to develop a gravity force of
thousands of pounds. Observing the following tips will ensure safe and efficient operation.
WARNING
Handle body fluids with care because they can transmit disease. No known test
offers complete assurance that body fluids are free of micro-organisms. Some
of the most virulent—Hepatitis (B and C) and HIV (I–V) viruses, atypical
mycobacterium, and certain systemic fungi—further emphasize the need for
aerosol protection.
D-6 PN JR-IM-10AG
APPENDIX E
References
List of References
Documents listed below* can be obtained by calling Beckman Coulter Customer Service at 1-800-742-2345
(U.S.A. or Canada) or by contacting your local Beckman Coulter office. They are also available at
www.beckmancoulter.com.
IN-181 How to use Quick-Seal Tubes with the Beckman Coulter Cordless Tube Topper.
IN-192 Use and Care of Centrifuge Tubes and Bottles
* For detailed information on a rotor, see the applicable individual rotor manual.
PN JR-IM-10AG E-1
References
List of References
E-2 PN JR-IM-10AG
Glossary
PN JR-IM-10AG Glossary-1
Glossary
Granulocytes — Generic name for three leukocyte Noryl — Modified thermoplastic polyphenylene
(white blood cell) types characterized by having oxide (PPO) used for floating spacers (part of the
granules in their cytoplasm g-Max system) and some polycarbonate bottle
caps (Noryl is a registered trademark of
Hard spin — Centrifugation run (5 to 7 minutes), GE Plastics.)
at high g-forces (4000 to 5000 × g) at ambient
temperature or at 4°C, used to separate fresh Pelleting — A centrifugal separation process in
plasma from cellular components which particles in a sample sediment to the
bottom of the tube (differential separation);
HDPE — High density polyethylene used for differential pelleting separates particles of
adapters different sizes by successive centrifugation steps
of progressively higher g force and/or longer run
Isopycnic — A method of particle separation or
duration
isolation based on particle buoyant density;
particles are centrifuged until they reach a point PET — polyethylene terephthalate used in some
in the gradient where the density of the particle adapters
is the same as the density of the gradient at
that point Plasma — Major component of blood made up
primarily of water, with substances such as
LDPE — Low density polyethylene used for tubes albumin, globulins, coagulation factors, and
and bottles electrolytes; distributes nutrients to the body,
absorbs and carries away waste products
Leukopheresis — Procedure in which leukocytes,
or white blood cells, are separated from whole Plasmapheresis — Procedure in which whole
blood blood is collected, platelets are separated via
centrifugation, and platelet-poor red blood cells
Leukocytes — See WBC (white blood cells)
are returned to the donor; plasma is returned to
Lymphocyte — A type of leukocyte formed in the the donor or collected for fractionation into
lymph nodes, other lymphoid tissue, and bone clotting factors and albumin
marrow; about a quarter of the white blood cells
Plateletpheresis — Procedure in which a unit of
in the circulating blood are lymphocytes
blood is taken to obtain plasma; following blood
Maximum volume — The maximum volume at separation, red cells are immediately reinfused
which a tube should be filled for centrifugation to the donor
(sometimes referred to as maximum fill volume
Platelets — Blood component responsible for
or nominal fill volume)
blood coagulation
Meniscus — The curved upper surface of a liquid
Polypropylene — Random block copolymer of
column that is concave when the container walls
ethylene and propylene used for certain tubes.
are wetted by the liquid and convex when they
are not Rack-type rotor — A rotor in which tubes are
placed in gamma-counter racks; the racks are
NaCl — Sodium chloride; a lower-density salt than
loaded into special plastic trays, which are then
CsCl, primarily used in lipoprotein type
loaded into carriers that swing up to the
separations
horizontal position during centrifugation
Neoprene — Black synthetic elastomer used for
Radel — Polyphenylsulfone (PPSU) used in plugs,
O-rings in some tube caps and bottle
cap closures, cannisters and other accessories
cap assemblies; should be used at temperatures
between –54 and 121°C (–65 and 250°F) Rate zonal — A method of particle separation,
based on differential rate of sedimentation, using
a preformed gradient with the sample layered as
a zone on top of the gradient
Glossary-2 PN JR-IM-10AG
Glossary
RBC — Red blood cells, or erythrocytes, carry Soft spin — Short centrifugation run (3 to
oxygen to the tissues and carbon dioxide to the 5 minutes), at low g-forces (2000 to 3000 × g) at
lungs for exhalation ambient temperature, used to keep small cells or
platelets in suspension while the larger cellular
RCF — Relative centrifugal field; the ratio of the components sediment; used to obtain platelet-
centrifugal acceleration at a specified radius and rich plasma and red blood cell concentrate from
speed (rω 2) to the standard acceleration of whole blood
gravity (g) according to the following equation:
Solution 555 — Beckman Coulter concentrated
rω 2 rotor cleaning solution; recommended because it
RCF = ---------
g is a mild solution that has been tested and found
effective and safe for Beckman Coulter rotors
where r is the radius in millimeters, ω is the and accessories
angular velocity in radians per second
(2π RPM/60), and g is the standard acceleration Spinkote — Beckman Coulter lubricant for metal-
of gravity (9807 mm/s2). Thus the relationship to-metal contacts
between RCF and RPM is:
Sucrose — A sugar (not a self-forming gradient)
2 used in rate zonal separations; generally used in
RCF = 1.12r ⎛------------- ⎞
RPM
⎝ 1000 ⎠ separating RNA, subcellular organelles, and cell
membranes
rmax — (Maximum radius) the position of the liquid
in the tube at the maximum distance from the Supernatant — The liquid above the sedimented
axis of rotation when the rotor is at speed material following centrifugation
rmin — (Minimum radius) the position of the liquid Svedberg unit, S — A unit of sedimentation
velocity:
in the tube at the minimum distance from the
axis of rotation when the rotor is at speed 1 S = 10-13 seconds
PN JR-IM-10AG Glossary-3
Glossary
Glossary-4 PN JR-IM-10AG
Beckman Coulter, Inc.
J Series Rotor Warranty
Subject to the conditions specified below and the warranty clause of the Beckman Coulter, Inc., terms and conditions of
sale in effect at the time of sale, Beckman Coulter, Inc. agrees to correct either by repair, or, at its election, by replacement,
any defects of material or workmanship which develop within seven (7) years after delivery of a J series rotor to the
original buyer by Beckman Coulter, Inc. or by an authorized representative, provided that investigation and factory
inspection by Beckman Coulter, Inc. discloses that such defect developed under normal and proper use. Should a
Beckman Coulter centrifuge be damaged due to a failure of a rotor covered by this warranty, Beckman Coulter will supply
free of charge all centrifuge parts required for repair.
Replacement
Any product claimed to be defective must, if requested by Beckman Coulter, Inc., be returned to the factory,
transportation charges prepaid, and will be returned to Buyer with the transportation charges collect unless the product
is found to be defective, in which case Beckman Coulter, Inc. will pay all transportation charges.
A defective rotor will be replaced by Beckman Coulter, Inc. at its then current list price less a credit based upon the age
of the rotor (years since date of purchase). The Buyer shall not receive credit until the claimed defective rotor is returned
to Beckman Coulter’s Indianapolis, Indiana, facility or delivered to a Beckman Field Service representative.
The replacement price (cost to Buyer) for the respective rotor shall be calculated as follows:
years
Replacement price = Current rotor list price × --------------
7
Conditions
1. Except as otherwise specifically provided herein, this warranty covers the rotor only and Beckman Coulter, Inc. shall
not be liable for damage to accessories or ancillary supplies including but not limited to (i) tubes, (ii) tube caps,
(iii) tube adapters, or (iv) tube contents.
2. This warranty is void if the rotor has been subjected to customer misuse such as operation or maintenance contrary
to the instructions in the Beckman Coulter rotor or centrifuge manual.
3. This warranty is void if the rotor is operated with a rotor drive unit or in a centrifuge unmatched to the rotor
characteristics, or is operated in a Beckman Coulter centrifuge that has been improperly disassembled, repaired, or
modified.
4. Each bucket, whether purchased with a rotor assembly or purchased separately, is covered by this warranty for
seven (7) years from the date of purchase, and will be replaced or repaired during such period according to the terms
and conditions of this warranty. The date of manufacture marked on the bucket may be earlier than the date of
purchase, and the expiration date marked on the bucket, which is seven (7) years after the date of purchase, may be
correspondingly offset.
5. Buckets should not be used after the expiration date marked on the bucket. If at the time of purchase the marked
expiration date is less than 7 years from the date of purchase, the expiration date becomes the date of purchase plus
seven (7) years. Use of a bucket after such expiration date voids Beckman Coulter’s warranty obligations with respect
to any rotor and/or centrifuge in which such a bucket is used.
Disclaimer
IT IS EXPRESSLY AGREED THAT THE ABOVE WARRANTY SHALL BE IN LIEU OF ALL WARRANTIES OF FITNESS AND OF
THE WARRANTY OF MERCHANTABILITY AND BECKMAN COULTER, INC. SHALL HAVE NO LIABILITY FOR SPECIAL OR
CONSEQUENTIAL DAMAGES OF ANY KIND WHATSOEVER ARISING OUT OF THE MANUFACTURE, USE, SALE, HANDLING,
REPAIR, MAINTENANCE, OR REPLACEMENT OF THE PRODUCT.
PN JR-IM-10AG Warranty-1
Beckman Coulter, Inc. J Series Rotor Warranty
Warranty-2 PN JR-IM-10AG
Related Documents
Rotors and Tubes CD (369668) Additional References
• Rotors and Tubes for Tabletop Preparative • Chemical Resistances for Beckman Coulter
Ultracentrifuges Centrifugation Products (IN-175)
• Rotors and Tubes for J2, J6, Avanti J Series • Beckman Coulter High Performance, High
Centrifuges Speed, High Capacity Rotors, Tubes &
• Rotors and Tubes for Preparative Accessories catalog (BR-8102)
Ultracentrifuges • HarvestLine System Liner Kit 369264
• Rotor Safety Bulletin (J-TB-093)
• Chemical Resistances for Beckman Coulter • Use and Care of Centrifuge Tubes and
Centrifugation Products Bottles (IN-192)
Included with shipment of instrument. • The JE-6B Elutriation System and Rotor
(JE6B-IM-9)
• The JE 5.0- Elutriation System (JE5-IM-13)
• Run Speeds for Stainless Steel Tubes
(L5-TB-072)
• How to Use the Beckman Coulter Cordless
Tube Topper with Quick-Seal Tubes (IN-181)
• Beckman Coulter Fraction Recovery System
(L5-TB-081)
• JCF-Z Zonal and Continuous Flow Rotor
(JCFZ-IM-12)
• Instructions for Using Micro Plus Multiwell
Plate Carriers (GS6-TB-011)
• Instructions for Using Microplate Carriers
(J6-TB-009)
www.beckmancoulter.com