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Medical Laboratory Instrumentation

Presentation · May 2014

DOI: 10.13140/RG.2.2.31677.67045

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Medical Laboratory
Instrumentation
‫أجهزة المعامل الطبية‬

Dr. Moustafa Moustafa Mohamed

Ahmed
Biophysics Department, Medical Research
Institute, Alexandria University
 Medical laboratory equipment provide a
means of measuring required substance
and metabolic waste products in urine and
blood.
 Blood Test
Blood cell tests include:
1- Red and white blood cell count
2- Platelets count
3- Hematocrite (Ht)
4- Mean cell volume (MCV)
5- Mean cell hemoglobin (MCH)
6- Mean cell hemoglobin concentration
(MCHC)
7- Blood chemical tests (acidity- pH
normally 7.36 to 7.44)
 Red blood cell count White blood cell count

Manually Automatically Manually Automatically

(100 to 1 dilution) (blood cell analyzer) (10 to 1 dilution) (blood cell analyzer)

Platelet cell count

Automatically
(blood cell analyzer)
 Hematocrit (Ht)

Percentage of solid part of  Plasma

blood
55%

WBC

RBC
45%
 Mean cell volume (MCV)

Mean cell volume (MCV):

MCV = Ht/ RBC count
 Mean cell hemoglobin (MCH)

Mean cell hemoglobin ( ‫متوسط حجم هيموجلوبين الخلية‬ 

: )(MCH)

 MCH = ( hemoglobin in grams/100 ml)/RBC

count
 Mean cell hemoglobin
concentration (MCHC)
 Mean cell hemoglobin concentration
(MCHC)

 MCHC= (Hemoglobin in grams at

100ml)/ Ht
 Blood Chemical Tests
 Check for:
- pH : normally 7.36 to 7.44 (slightly alkaline).
- Glucose: lactic acid, lactose sugar.
- Non-protein nitrogen substances: amino
acids, peptides, urea waste, and uric acid
- Lipids: fatty acids of cholesterol and
triglycerides
- Proteins: plasma, albumin, globulins, and
fibrinogen
- Enzymes
Blood serological tests
 Testing for agglutination (clamping) or cells
due to the addition of antigens
 Blood bacteriological test: include growth of
blood bacteria in a Petri dish.
 Histological tests: studies of small thin tissues
samples under microscope. Specimen are
obtained by cutting tissues with a precision
slicer known as microtome.
 Equipment
 Centrifuges
 Suction devices
 Colorimeters
 Spectrophotometers
 Blood/cell and gas analyzers
 Chromatographs
 Autoanlyzers
 Computer-based record and operation system
 Flame, Colour meter, and
spectrophotometer
 Colour meter
 Instrument for measuring concentration of
substance by measuring its colour .
 Flame photometer
‫فوتومتر اللهب‬
 Instrument for measuring the
concentration of any easy ionized element
by measuring the intensity of its emitted
light when excited by flame
Spectrophotometer
‫االسبكتروفوتومتر‬
Instrument for measuring the
concentrations of easy ionized
elements by measuring the intensity
of its emitted light spectrum when
excited by flame
Absorption Spectroscopy

Sample

h h
M
M M M
M* M* M *
in M
M* out
M

reflection scattering
Absorption Spectroscopy
Generic Instrument

P0 P

Light Mono-
Sample Detector
Source chromator

P0 = intensity of light into sample

P = intensity of light out of sample
Absorption Spectroscopy

 The more photons the sample absorbs, the

lower the intensity (transmission) at the
detector.
 Transmittance (T)
P
T
 0T1 P0
 T is independent of P0
 %T = T x 100
 Absorption Spectroscopy

 sample with T = 50%

P0 = 1.00 P = 0.50 P = 0.25 P = 0.125

Absorption Spectroscopy
1
T

0.5

0
Concentration or Path Length
Absorption Spectroscopy

 Absorbance (A)
P0 1
A  log  log   log T
P T
%T
T ; A  2  log %T
100
0A
Absorption Spectroscopy
A

Concentration or Path Length

Beer’s Law

A = bc
Beer’s Law
 Light sources

 Different light sources for different regions of the

spectrum
 UV/Vis
 Tungsten Lamp 320-2,500 nm - run an
electrical current through a wire in vacuum
 Deuterium arc lamp, 200-400 nm - electrical
discharge in D2
 Laser source
 Monochromators
 Mono - one; chromatic - color
 Prisms and gratings - disperse (spread out) light
according to wavelength

h

Prism
 Monochromators
 n = d(sin I + sin r)
 I = constant; therefore   r

r I

d
 Sample Holders
 Cuvettes
 flat surface best - better reproducibility
 avoid fingerprints, dust, etc. on surface
 must be transparent in region of interest
 Sample Holders

 UV - quartz
 Visible - glass, quartz
 IR - NaCl
 Detectors

 Detector converts incident light to an

electrical signal that we can measure and
process
 Detectors

 Ideal Characteristics
 sensitive
 linear
 flat response v. 
 stable
 fast
Detectors - photo tube

 Good for UV and Visible

h

e-
amplifier
-V
dynode
 Detectors - photo multiplier
 Characteristics
 sensitive (single photons)
 linear
 flat response v.  within limitations
 stable w/ time (sensitive decreases over time,
weeks to months)
 fast
Detectors - Array
 Channel plate (similar to multiplier)
 photodiode array (less sensitive than
multiplier)
 charge coupled device (CCD) (more
sensitive than multiplier)
 Do not need a monochromator
 Color-meter
 An optical electronic device that
measures the color concentration of a
substance in solution.
 Optical color filters are used to select a
narrow wavelength.
 Basic colorimeter analysis involves the
precise measurement of light intensity.
 Transmittance is defined as:
I1= T * I0
I0 = Initial light intensity and I1= attenuated light
intensity
 The results are displayed in percent optical color
transmittance or absorption to indicate
hemoglobin concentration.
I1
T  x100 percent
Io
 Single Beam Instrument
Slit

Light Sample Detector

Source Filter,
(Tungsten Mono- Quartz Photo-
Lamp) Chromator, Cuvette multiplier
Grating
 Double Beam Instrument
Beam Chopper Semi-transparent
Mirror

Sample

Tungsten Slit Quartz Photo-

Grating
Lamp Cuvette multiplier

Reference
Mirror Mirror

Mirror
Color meter- filter photometer
 The concentration of the unknown solution
can be found from the following
relationship:
Cu= Cs Au/As
where
Cu= unknown concentration
Cs= standard concentration (for calibration)
Au= unknown absorbance
As= standard absorbance
 Light passes through an optical color
filter, is focused by lenses on the
reference and sample cuvettes and
falls on the reference and sample
photodetectors.
 The difference in voltage between the
two detectors is increased by a dc
amplifier and applied to a meter.
A calibration procedure is as
follows:
1- Ground the amplifier input (V1) and adjust
potentiometer (R1) for 0 volt
2- Remove the ground and place reference
concentration in cuvettes 1 an 2
3- Adjust potentiometer R1 for 0 V
4- Leave the reference concentration in cuvette 1
and replace cuvette 2 with a cuvette containing
the sample
5-Read the unbalanced voltage on the meter in
percent transmittance or absorbance units.
 Spectrophotometer
 The spectrophotometer measures light
absorption by a liquid substance at various
wavelengths.
 The components of an unknown material
can be determined or the concentration of a
number of known substances can be
measured.
 A monochrometer uses a diffraction grating or
prism to disperse the light from the lamp (slit S1).
 The light is broken into its spectral components as
it arises from slit S2 and falls on the sample in the
cuvette,
 The angle of the diffraction grating determines
light wavelength.
 The mirror reduces equipment size.

 light output, photodetector sensitivity, and
sample substance absorbance changes with
wavelength, and this necessitates zero
calibration for each wavelength
measurement.
 The double-beam spectrophotometer
accomplishes this automatically by beam path
switching (sample to reference) via a
mechanical shutter or rotating mirror.
Maintenance
 Maintenance includes:
 Calibration adjustment and replacement
of light source bulbs and photo
detectors.
 Mechanically rotating assembles
(mirrors, diffraction grating) will
occasionally malfunction.
 The electronics is very reliable.
Atomic Spectroscopy
AE measures photons emitted from excited 

(by heat) sample

A
A*
A*
A
A Detector
monochromator
A
A
A
Atomizer/vaporizer
 Flame Photometer
 The flame photometer measures the color
intensity of flame that supported by oxygen
and specific substance.
 The basic schematic shows that a reference
gas containing a lithium salt causes a red
color to shine on the reference photo
detector through the reference optical filter.
 A yellow or violet light from sample sodium
or potassium falls on the sample photo
detector.
 Flames
Flame

Atomizer Fuel
Oxidant

Aspirate sample
into flame Sample
 Atomizers - Flames
 Use different fuel/oxidant combinations to
get different temperatures
 C2H2/Air : 2400 - 2700 K
 C2H2/O2 : 2900 - 3100 K
Flame photometer-simplified
schematic
Calibration
 Basically, the flame photometer is
calibrated in a manner similar to the
colorimeter
 Continuous calibration can be
accomplished by inspiration of air and
lithium.
 the output is read in units of sodium or
potassium concentration.
Maintenance
 Maintenance includes calibration, adjustment
and replacement of light bulbs and photo
detectors.
 Aspiration devices and flame chambers
require cleaning.
 Also mechanical rotating assembles (mirrors,
diffraction grating) occasionally malfunction
 Electronic failures are usually infrequent.
 Sensor terminology
Sensitivity
Defined as the slope of the output
characteristic curve,
or
An output voltage change for a given change
in input parameter.
Detection Limit
The minimum input of physical 

parameter that will create a detectable

output change,
Sensitivity error
The sensitivity error is the departure 

from the ideal slope of the characteristic

curve.
Ideal curve and sensitivity
error
Typical pH-electrode characteristic-curve
showing temperature sensitivity.
Dynamic range
The dynamic range is the total range of 

the sensor from minimum to maximum.

Precision
The degree of reproducibility of a 
measurement
Resolution
The smallest detectable increment range of 

input parameter that can be detected in the

output signal.

Accuracy
The maximum difference that will exist 

between the actual value and the indicated

value at the output of the sensor
offset
The output that will exist when it should be 
zero.
Linearity
Actual measured curve of a sensor departs
from the ideal curve
Din max .
x100 Nonlinearity %=
IN f .s.
Din (max) is the maximum input deviation
INf.s. is the maximum, full-scale input
Ideal versus measured curve
showing linearity error
Hysteresis
A transducer should be capable of 

following the changes of the input

parameter regardless of which direction
the changes is made; hysteresis is the
measure of this property
Hysteresis curve
F(x)

F(x)2

F(x)1

x
B
 Response Time
The response time can be defined as 

the time required for a sensor output to

change from its previous state to a final
settled value within a tolerance band of
the correct
V
new value
V

T
T
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