LAMBDA MINIFOR laboratory fermentor

The LAMBDA MINIFOR bench-top laboratory fermenter

and bioreactor was developed as a result of the need to
construct a small laboratory fermentor for volumes from
35ml to over 6 liters. Based on long personal practical
experience of fermentations we wanted to create a
fermentor, which was easy to use and with the
capacity to measure and control all the important
parameters of the biological culture.

The fermentor had to take up minimum space on the

bench but with good access to all parts. Several
fermentors should when placed side by side be suitable
for the optimization of the parameters of growth of
culture or optimization of bio-transformations etc.

Each fermentor should be able to work independently

or be connected to a PC for advanced regulation
and extensive data treatment.

To keep the cost of the MINIFOR fermentor-bioreactor low, without compromising

quality, several new ideas and innovations have been introduced:

• Instead of a fermentor flask with a stainless steel cover, which is expensive, we use
whole glass vessels with threaded fittings. They have been used for many years in
cell culture and are proved to maintain perfect sterility. Thanks to this concept, the
MINIFOR fermenter-bioreactor is set up in shortest possible time.

• Instead of a traditional propeller agitator, which requires an expensive motor and

magnetic coupling, we have introduced a new up-and-down agitation. A motor
together with an inexpensive membrane perfectly assure sterility and produce an
efficient mixing without formation of a vortex (no baffles needed). At the same time
this type of mixing is gentler on cells and produces less foam. Novel biomimicking
“fish-tail” stirring discs offer maximum mixing efficiency without cutting edges.

• The culture is heated by heat radiation produced in a parabolic radiator with a gold
reflector placed under the fermentation vessel. The heat is absorbed gently in the
culture similarly to the sun heating water. There is no overheating of the culture at
any volume and expensive double-wall vessels with thermostatic baths are eliminated.
At the same time tubing and cables disappear making the fermentor less complex.

• As far as possible expensive pieces of equipment have been replaced by new high
performance plastics.

• By using modern microprocessors it has been possible to place all the electronics in
the front part of the instrument. This makes the fermentor unbelievably compact and
eliminates the casing tower. Despite its small size six parameters are measured and
controlled in the basic configuration of the MINIFOR.

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Technical Description
Basic unit

The main feature of the MINIFOR is that all the electronics, power
supply, IR heater, air valve, mass flow meter, cables and tubing are
integrated in one base unit, which is used as the support for the
fermentation vessel and other accessories. The platform
arrangement of the base unit makes all parts of the fermentor
clearly visible and easily accessible from all sides despite base
dimensions of only 22 x 40cm (approx. an A4 sheet of paper).

Up to five reagent bottles in magnetic holders can be placed behind the fermentor vessel
and up to 4 pumps can be placed on adjustable holders mounted on bars at the rear part of
the base unit. The sockets are located on the rear side of the base unit thus removing cables
from the work area.

Measurement and regulation

The control panel consists of an LCD display and control

buttons. All parameters (temperature, pH, pO2, air flow
rate, agitation and one free selectable parameter, e.g.
pCO2, optical density, antifoam etc.) are visible at a
glance on a large LCD back light display (4 x 40 digits).
The controls are simple and logical.

On most parameters the limits of low or/and high alarm can be set. After alarm activation an
acoustic signal is heard, the indication appears on the display and an electric signal is
generated on the rear side connector of the fermentor. Each fermentor can be operated in an
autonomous way or can be coupled to a PC (over RS 485) using a special fermentation
program FNet (for up to 6 fermentors in parallel) or SIAM software. The latter allows almost
unlimited possibilities for control and data processing.

Fermentation vessel

The standard fermentation vessel has a working volume of up to 1.7 liter

(other vessels with working volumes from 35 ml to up to 6.5 liters can also be
provided). The vessels are made of high quality Pyrex glass with one large
threaded central opening for the fixation of the membrane and vibro-mixer
and 6 to 8 threaded necks for the connection of different probes, air outlet,
sample withdrawal, inoculation etc. An adjustable holder maintains the vessel
in position. The fermentation vessel is sterilized in an autoclave.

Vessel type: 0.3 0.4 1 3 7

Volume [L]: 0.035 – 0.4 0.15 – 0.45 0.3 – 1.7 0.5 – 3 1 – 6.5
Height [cm] 34 22 34 37 50
Diameter [cm] 22 23 25 34 30
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Dimensions

A lot of effort was made to make the MINIFOR as small and as compact as possible. The
platform arrangement of the fermentor makes an easy control and access to all parts from
all sides. The distance between the reagent bottles, pumps and the fermentor vessel is the
shortest possible.

Agitation

Instead of a traditional propeller agitator a new vibromixer is

used in the MINIFOR. A strong motor moves one or more
stirring discs up and down. The major advantage is an
efficient mixing and aeration of the culture medium together
with very complete separation of the inside of the vessel from
the outside by a low cost silicone membrane. No vortex is
built up and baffles are eliminated. This type of agitation is
also gentler on the cells and foaming is reduced. A very
efficient mixing is achieved with the novel elastic “fish-tail”
stirring discs. They combine maximum mixing efficiency
without cutting edges - the optimal choice for cell cultures.
The frequency of agitation is controlled by a microprocessor
and can be varied through a broad range. The input of air is
made through elastic self-cleaning micro-sparger.

Temperature control

A new infrared (IR) radiator with a gilded parabolic reflector

is used to warm the culture broth. The heat radiation (150W)
is concentrated on the bottom of the vessel where it is
absorbed by the medium in a similar way to the sun heating
water. There is no overheating at any volume of the culture
(common with heaters placed directly in the medium). Thanks
to the low heat capacity of the IR source, overshooting of the
temperature is reduced and the temperature can be
controlled more precisely. The temperature sensor is
placed directly in the pH sensor and is used at the same
time for an automatic correction of pH and pO2 electrodes.

pH measurement and control

pH is measured by a combined, sterilizable pH electrode

with incorporated temperature sensor. Thanks to the
multiple Variopin plug it can be sterilized without any protection. The two-point calibration of
the electrode is semiautomatic. The pH value has an automatic temperature correction.
The addition of acid or base is controlled by a microprocessor. The flow rate of the
peristaltic pumps PRECIFLOW, MULTIFLOW, HIFLOW or MAXIFLOW is varied between
0 to 100%, which makes the pH control smoother than with the common ON/OFF switching
of fixed speed pumps. The unique PUMP FLOW INTEGRATOR (option) when coupled to the
pump enables monitoring of the pump’s activity during the process. This yields precise kinetic
data concerning the culture state and activity.

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pO2 measurement and regulation

A sterilizable Clark-type electrode with large cathode

measures the concentration of dissolved oxygen with a glass
reinforced TEFLON membrane giving fast response times and short polarization. The major
part of the membrane is protected against mechanical injury by a thin wall of PEEK. The
microprocessor performs a semiautomatic two-point calibration with automatic temperature
compensation. The regulation of dissolved oxygen is obtained by a variation of the flow
rate of air.

Air input

The flow rate can be set from 0 to 5 l/min in 0.1 l/min steps. A precise mass-
flow meter is used. This measurement is independent of the pressure and
temperature variation of the air. Commonly used floating ball capillaries
(rotameters) give inaccurate readings in this case. A proprietary proportional
air valve adjusts the flow rate continuously.

A new self-cleaning elastic air micro-sparger has been developed. Its special construction
allows an automatic elimination of salt deposits, which would block the airflow in normal
spargers. This is important particularly for micro-spargers having very narrow channels.

Air output

Used air is cooled on a glass condenser and filtered by a PTFE (Teflon) filter. An optional
Peltier condenser, which does not require cooling water, can be used to remove water
vapor from the air stream.

Inoculation and sampling ports

Inoculation, addition of acid or base and sample removal is made through

four stainless steel capillaries equipped with LAMBDA double seal PEEK
fittings.

High-quality peristaltic pumps

Up to four high-quality peristaltic pumps PRECIFLOW, MULTIFLOW,

HIFLOW or MAXIFLOW can be placed on holders fixed on two bars at
the rear of the fermentor. They are connected by a single cable to the
sockets on the rear side of the fermentor. Since the pumps are not
integrated into the fermentor they can be used for other applications
elsewhere in the laboratory (e.g. for chromatography etc.). This
represents considerable savings for the user.

A new connection system provides double sealing of the tubing and, therefore, reduces
strongly the contamination probability during the transfer of solutions into the vessel.

4
Gas flow controller

LAMBDA MASSFLOW is a new mass flow controller system specially

designed for the use together with LAMBDA laboratory bioreactors and
fermentors. It allows the control of pH of cell culture by controlled addition
of gaseous CO2, control of nitrogen or of any other gas with suitable
controller. However, it can be also used quite independently, since all
functions can be accessed from the front panel of the MASSFLOW.

A high quality laminar mass flow sensor measures the flow rate. The result appears on the
digital display. The mass flow cell has a very low pressure drop and a linearity error less than
±3% reading. The repeatability is better than ±0.5% reading. The flow rate is regulated by a
special proprietary proportional needle valve controlled by a microprocessor. The flow
rate can be programmed and volume totalized. MASSFLOW allows a precise, automatic
control of pH in cell cultures without need of any other gas station.

Continuous fermentation (chemostat)

A scale adaptor (optional) allows weighing of the fermentor.

It is simply placed under the front part of the fermentor body
and connected to the X-channel input of the fermentor. By
means of a pump connected to the ”Pump X”-socket of the
fermentor, the weight (volume) of the culture can be kept
constant. This function is included in the start-up kit already.
This allows the running of continuous cultures at low cost.

PC software

FNet was specially developed for the monitoring and

control of fermentation processes and cell cultures with
the LAMBDA MINIFOR fermentor. The software runs under
Windows NT, 2000, XP, Vista and Windows 7:

• Easy to install and to use

• No need of programming knowledge
• Recognizes the connected fermenters at start-up
• For up to 6 fermentors, 12 integrators and 6 pumps on one PC
• All the cables use standard connectors

For very high requirements we propose an industrial fermentation software SIAM. There
are almost no wishes, which could not be satisfied with SIAM. Special functions can be added
according to the user’s needs.

5
Technical parameters
The LAMBDA MINIFOR fermentor/bioreactor is controlled by two microprocessors.

Power: Mains 100-240 V AC/50-60Hz, 510W, CE conform

Dimensions: 22x40x38cm (WxDxH)
Display: LCD 4 x 40 digits with backlight illumination
Fermentor vessel: Pyrex glass with 6 to 8 side necks 0.3, 0.4, 1, 3 and 7 l
Working volumes: 35ml to over 6 liters
Temperature control: IR radiation heat source with gilded reflector 150W
Regulation: from 5°C above RT to over 70°C
Measurement: from 0 to 99.9°C in 0.1°C steps
Precision: ± 0.2°C (0 to 60°C)
Sensor: Pt 100 incorporated in the pH sensor
pH control: sterilizable pH electrode (Mettler), pH 0-13, with automatic
temperature correction, Variopin connector, two-point
semiautomatic calibration
Resolution: 0.01 pH unit
Precision: ± 0.02 pH unit
Pumps: up to 4 independent peristaltic pumps (PRECIFLOW,
MULTIFLOW, HIFLOW or MAXIFLOW) with speed variation
from 0 to 100 % can be used with MINIFOR
pO2 control: sterilizable Clark type oxygen sensor, PEEK body, Mettler
membrane module, automatic temperature correction
Range: 0 to 25 mg oxygen/l, in 0.1 mg/l steps, automatic control
through regulation of air flow rate
Air flow: 0 to 5 l/min. in 0.1 l/min. steps, mass flow meter, linearity
± 3%,
Reproducibility: ± 0.5%
Control: proportional valve
for supplied air pressure: between 0.05 – 0.2 MPa (max.)
Agitation: 50 W Vibromixer 0 to 20 Hz (0 to 1200 rpm), in 0.1 Hz
steps, with 1 or more stirring discs
Sterility comparable to magnetic coupling
Selectable parameter: an additional parameter can be controlled by the
instrument (foaming control, pCO2, optical density,
conductivity, etc.)
Sampling/addition ports: up to 4 stainless steel needles with LAMBDA PEEK
double-seal fittings can be used for sampling and addition
of correction solutions
Working temperature: 0 - 40 °C
Working relative humidity: 0 - 90 % RH (non condensing)
Security: conform to IEC 1010/1
Weight: 7.5 kg
PC control: complete PC control and data treatment using FNet or SIAM
fermentation software (optional)
Warranty: 2 years

LAMBDA Laboratory Instruments LAMBDA CZ s.r.o.

Dr. Pavel Lehky Lozibky 1
Imfeldsteig 12 CZ-61400 Brno
CH-8037 Zürich Czech Republic
Switzerland
Tel: +41 (0)44 450 20 71 Tel/Fax: +420 545 578 643
Fax: +41 (0)44 450 20 72 Hotline: +420 603 274 677
E-mail: [email protected]
Web: www.lambda-instruments.com www.bioreactors.eu