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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 35 ml 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• As far as possible expensive pieces of equipment have been replaced by new high performance plastics.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.
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 × 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.
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 × 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.
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 5 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.
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.
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.
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 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.
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.
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.
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, addition of acid or base and sample removal is made through four stainless steel capillaries equipped with LAMBDA double seal PEEK fittings.
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.
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.
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.
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.
The LAMBDA MINIFOR fermentor/bioreactor is controlled by two microprocessors.
|Power:||Mains 100-240 V AC/50-60Hz, 510W, CE conform|
|Dimensions:||22 × 40 × 38 cm (W × D × H)|
|Display:||LCD 4 × 40 digits with backlight illumination|
|Fermentor vessel:||Pyrex glass with 5 to 8 side necks 0.3, 0.4, 1, 3 and 7 l|
|Working volumes:||35 ml to over 6 liters|
|Temperature control:||IR radiation heat source with gilded reflector 150 W|
|Regulation:||from 5°C over RT to 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%|
|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|
|PC control:||complete PC control and data treatment using FNet or SIAM fermentation software (optional)|