pbr101 photo bioreactor guide to setup and operationspbr101 photo bioreactor guide to setup and...
TRANSCRIPT
PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
Table of Contents
1. Safety ............................................................................................................................... 2
2. Unpacking and Parts Identification .................................................................................. 4
3. General Description and Specifications ........................................................................... 7
4. PBR101 Installation and Setup ....................................................................................... 10
4.1. Operating Environment Requirements .................................................................. 10
4.2. Connecting the PBR101 to Power and a Network .................................................. 10
4.3. Setting up the Jacket Assembly .............................................................................. 13
4.4. Preparing the Culture Vessel Assembly for an Experiment .................................... 17
4.5. Attaching pH and Reference Electrode, and Culture Temperature Probe ............. 23
4.6. Connecting the LED Assembly Power Cables ......................................................... 26
4.7. Connecting Gas to the PBR101 and Culture Vessel ................................................ 27
4.8. Powering on the PBR101 ........................................................................................ 29
5. Setup and Operation of the TP2501 Turbidistat Pump (optional) ................................ 30
5.1. Key Features of the TP2501 .................................................................................... 30
5.2. Parts Identification ................................................................................................. 30
5.3. Assembly ................................................................................................................. 32
5.4. TP2501 On-Board Operational Controls ................................................................. 37
6. Maintenance Procedures ............................................................................................... 38
7. Consumables and Field Serviceable Parts ..................................................................... 41
8. Troubleshooting and Technical Support ........................................................................ 42
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
1. Safety
General Safety Guidelines. Please read the following carefully before
attempting to operate the PBR101 or related equipment.
• This manual contains important information and warnings which must be followed in
order to ensure safe installation and operation.
• The manufacturer is not responsible for any damage or injury caused by improper setup
or operation.
• Remove all packaging and protective materials before setting up and connecting the
PBR101 to the power supply.
• Use only components supplied by the manufacturer.
• Keep the outside of the device dry and avoid working in a high humidity environment.
• Water and other liquids should only be placed in vessels designed for this purpose and
according to instructions included in this manual.
• Before switching on the equipment, make sure that it is set to the proper line voltage.
Risk of Electrical Shock. You must follow these general electrical
safety guidelines.
• Perform a routine check of all devices and their wiring.
• Replace any worn or damaged electrical connectors and cords immediately.
• Use appropriate electrical extension cords/power bars and do not overload them.
• Install the device on a stable surface such as a laboratory bench. Keep away from wet
floors and counters.
• Do not touch the device, electrical sockets, outlets or switches if your hands are wet.
• Do not alter or modify any electrical parts of the device or its components.
• Any interruption of any protective conductor inside or outside the PBR101 or related
equipment or disconnection of the protective conductor terminal will make the device
dangerous. Intentional interruption is prohibited.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
• Capacitors inside a device may remain charged and could cause an electric shock hazard
even if it has been disconnected from all voltage sources.
• Any adjustment, maintenance and repair of the opened device under voltage must be
avoided as far as possible and, if inevitable, the activities must only be carried out by
qualified personnel.
• The use of makeshift fuses or short-circuiting of fuse holders or circuits is prohibited.
• It is not permitted to connect non-OEM equipment to the PBR101 or related devices as
this may lead to equipment damage, electrical shock, and/or fire.
Risk of overpressure of the gas lines and solenoid(s).
• The gas lines and solenoid may be damaged from over-pressure by inlet gas, potentially
resulting in operator injury and equipment damage.
• Do not apply a source gas pressure that exceeds 25 psig. Reduce the pressure of the
inlet gasses through a regulator or apply a pressure relief valve that is properly adjusted.
• It is highly recommend that regulators be used at each PBR101 for controlling the inlet
gas pressure, in addition to regulation at the source. This will also ensure proper gas
pressure when several PBR101s are fed from a common source, which can be
maintained at an adequate pressure.
Caution: Do not look directly at exposed LED lamps in operation.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
2. Unpacking and Parts Identification
The PBR101 is shipped in one large box which may contain smaller boxes, bags, and envelops.
The PBR101 components and accessories are packed using a large amount of packing materials
to ensure damage-free delivery. Unpack your PBR101 and save the packing materials for
possible future use.
Verify that you have received the correct items and that nothing is missing using Table 1 and
Figures 1 & 2. If any part of your order was damaged during shipping, is missing, or fails to
operate, please immediately contact Phenometrics Technical Support at:
+1-517-884-4361
Table 1: PBR101 components shown in Figures 1 and 2
Item Quantity Notes
a Control Tower 1
b Jacket Assembly 1
c LED Assembly 1
d Router 1 or more Size and number will vary depending
on the number of PBR101s ordered
e CD 1
Contains this Guide, Algal
CommandTM software program and
Algal Command manual
f Power Cable 1
g Ethernet Cable 1
h Culture Vessel 1 Port configuration may vary; see
section 4.4
i Jacket Temperature Probe 1 May be packed inside the Culture
Temperature Probe
j Stir Bar 1
k Turbidometer Sensor Cable 2
l Black O-Ring 1
m Glass Lens 1
n Red O-Ring 1
o Vessel Cap and Filter 1 Filter is important to prevent
microbial contamination of the vessel
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
p Tygon Tubing (12” & 24”) 2
q Luer Caps 4
r Luer Fittings, male 4
s Luer Fittings, female 4
t pH Electrode (optional) 1 White box labeled Pheno-003
u Reference Electrode (optional) 1 White box labeled Pheno-002
v Culture Temperature Probe 1 White box labeled Pheno-001
Figure 1: PBR101 Components (compare to Table 1)
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
Figure 2: Additional components (compare to Table 1)
If you ordered the optional Turbidistat Pump Module (cat. no.
TP2501), see Section 5 for additional parts, setup, and operating
instructions.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
3. General Description and Specifications
The Phenometrics PBR101 compact bench-top Photo Bioreactor was specifically designed for
the optimization and study of the growth of microalgae and cyanobacteria. It provides for a
great number of algal growth conditions to be controlled, monitored, and recorded by
computer, for accurate emulation of local environmental conditions, determination of optimal
growth conditions, or both. This allows for the optimization of an individual strain under given
production conditions, such as an outdoor raceway, or the selection of the best strain for use
under existing environmental conditions.
Virtually unlimited user-defined system control combinations are accomplished through Java
scripting in Algal Command, a customized software package written specifically for the PBR101.
The Java script of Algal Command is open-sourced, which allows advanced users to further
customize system control. Up to 256 separate PBR101 systems can be controlled from a single
laptop or desktop computer, simultaneously storing all of the data together. This allows for the
most rapid diversification of experimental conditions possible, resulting in the highest-possible
level of R & D or scientific study.
The combination of growth optimization and dramatically-reduced development times results
in a significant reduction in startup and operational costs (CAPEX) while at the same time
increases production yields with a reduced time-to-market (resulting in greatly enhanced
profit). This same increase in productivity can also be of equally significant value in a pure
research environment.
The following variables may be controlled using the PBR101 and Algal Command:
• 3000 μ-Einstein LED specially designed lighting system with conical vessel that
accurately mimics BOTH raceway or production tube bioreactors (controllable from 0-
3000 in steps of 25 μ-Einsteins.
• Fully programmable Diurnal Cycles; you set the length of sunlight day, and also the hour
of peak sunlight.
• Fully programmable heating and cooling, linkable to the diurnal cycle; you can
independently set the hour of peak heating (if different from peak sunlight).
• Active cooling and heating, from 5 - 50°C
• Algal Command Software provides for complete, simultaneous, integrated control of up
to 256 bioreactors from a single computer, with collation of all data. All the reactors are
linked together via Ethernet Hub.
• CO2 programmable and controlled from Algal Command.
• Mixing through active magnetic stirring and/or gas sparging.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
• Active sparging of up to two gasses
• Temperature measurement
• True Turbidostatic measurement
• Field-configurable, fully autoclavable, low cost durable polycarbonate reactor vessel
• The reactor vessel can have as many inlets and outlets as you require
• 100% (+/- 10%) directly-proportional (by volume) up-scale to pond, raceway, or indoor
reactor
• pH measured and controlled by Algal Command
• Custom-engineered Turbidostatic Pump allows for true turbidistatic operation; fully
programmable and computer-controlled.
Up to 256 PBR101s may be matrixed together, all
controlled simultaneously from a single computer
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
SPECIFICATION: DETAIL:
Culture vessel volume 700 ml
Working volume 150 – 600 ml
Light intensity Variable up to full ocean sunlight at the equator (>3000
mmol photons m-2 s-1) in steps of 25
Temperature control Active cooling and heating
Culture temperature range 10-50°C
Diurnal + Heating and Cooling
cycles Fully programmable and independent
Ability to control and measure pH 0-14; may be controlled automatically using CO2 gas
Mixing Magnetic stirrer or bubbling (sparging)
Gas Control Computerized; CO2 or other as required; optional second
gas inlet available
Growth rate measurement Continuous; via turbidistat
System configuration Customizable with specialized probes/configurations
Dimensions H-482mm, W-241mm, D-317.5mm, (H-19”, W-9.5”, D-12.5”)
Weight 7 kg (15lbs)
Power consumption 2.4 Amps
Voltage 110-120V/220-240V 50/60 Hz
Heat output 470 BTU/hr
Minimum PC requirements:
Windows 7 compatible computer, Mac OS 10.4, Linux,
minimum 2 GB memory, CD-ROM, Ethernet, Super VGA
display with a minimum 800 X 600 resolution and 256 colors
Sterilization Entire reactor vessel assembly, including culture medium,
may be autoclaved on the “Liquid” cycle
Turbidistatic pump True turbidistatic operation; fully programmable from
growth phase
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
4. PBR101 Installation and Setup
4.1 Operating Environment Requirements
Choose a suitable location for installing the PBR101 based on the follow operating
requirements and specifications of the instrument:
• Dimensions: H-482mm, W-241mm, D-317.5mm, (H-19”, W-9.5”, D-12.5”)
• Weight: 7 kg (15lbs)
• Power Consumption: 2.4A
• Voltage: 110-120V/220-240V 50/60 Hz
• Heat output: 470 BTU/hr
• Recommended spacing: allow a minimum of 12” around all sides of the instrument to
provide adequate ventilation and access.
• Recommended relative humidity not greater than 60%.
• Install on a flat, stable surface such as a laboratory bench.
4.2 Connecting the PBR101 to Power and a Network
• Connect the Power Cable (f) to the power supply on the side of the Control Tower (a) as
shown in Figure 3.
• Make certain that the red voltage switch, shown in Figure 3, is set the correct line
voltage. Failure to do so may result in permanent damage to your equipment, electric
shock, and fire.
• Connect the Power Cable (f) to a power source. It is recommended that the PBR101 be
connected to power through a properly sized surge protector to prevent electrical
damage in the event of fluctuations in the line voltage.
• Connect Ethernet Cable (g) to the PBR101 and the supplied Router (d) as shown in
Figure 4.
• Connect the Ethernet Cable (g) to your computer (not supplied) and appropriate port in
the Router (d).
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
Figure 3: Location of power supply on right side of Control Tower. Ensure the
red voltage switch is set to the proper line voltage (right panel).
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
Figure 4: Connection of Ethernet Cable (g) to Control Tower (a) and Router (d)
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
4.3 Setting up the Jacket Assembly
• Place the Jacket Assembly (b) in the square relief section of the PBR101 Control Tower
(a) table top, as shown in Figure 5. Notice that the wiring with the green plug is facing
toward the Control Tower (a).
Figure 5: Placement of the Jacket Assembly (b) on the
Control Tower (a) table top.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
• Connect the Jacket Assembly Control Connector (green plug) to the Temperature
Controller socket on the Control Tower (a) as shown in Figure 6. Align the plug as shown
and push firmly to fully seat the connector.
Figure 6: Jacket Assembly (b) control connector (green plug) shown plugged into
the corresponding Temperature controller socket on the Control Tower (a)
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PBR101 Photo Bioreactor
Guide to Setup and Operations
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• Connect a Turbidometer Sensor Cable (k) to the Emitter port on the Control Tower and
to the Emitter module. The Emitter module is facing the Control Tower (a). Upon close
inspection, you will notice it has the letter “E” molded into its cover as shown in Figure
7.
• Connect a Turbidometer Sensor Cable (k) to the Detector port on the Control Tower and
to the Detector module. The Detector Module has a “D” molded into its cover.
Figure 7: Identification of the Emitter (left panel) and Detector (right panel)
modules
Caution! Do not let any liquids contact the Emitter or Detector Modules.
Contact with liquids can damage these components and cause electric
shock and fire.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
• Carefully screw the Jacket Temperature Probe (i) into the Jacket Assembly (b) in the
threaded hole located in the top corner as shown in Figure 8. Plug the other end into
the “Jacket Temp” port on the Control Tower. When screwing the Jacket Temperature
Probe (i) into the Jacket Assembly (b) make sure that the cord can turn freely without
being twisted or kinked. Failure to do this carefully may result in damage to the
probe.
Figure 8: Connection of the Jacket Temperature Probe (i)
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
4.4 Preparing the Culture Vessel Assembly for an Experiment
The Culture Vessel ports and can be configured in a variety of ways. Generally, there needs to
be, at a minimum, an input for gas (usually CO2). An exhaust port is provided in the cap. This is
the standard configuration of the vessel supplied with the PBR101 (see Figure 11). Ports for gas
both input and exhaust must be properly filtered in order to avoid contamination of the
experimental culture. We recommend at least a 0.45 µm filter, one of which is supplied for
the cap exhaust port.
If you also purchased the TP2501 Turbidistat Pump, then the vessel (h) will also have three
additional ports as shown in Figure 11, for a total of four vessel ports.
In a typical experiment using the standard port configuration, follow these steps for completing
tubing connections to the ports and charging the vessel with media. The order of assembly for
the vessel cap assembly is shown in figure 9.
1. Clean the vessel using the following guidelines:
• Wash the vessel with only mild detergent and a soft cloth or bottle brush. Do not
use harsh laboratory detergents or scrubbers as these may scratch the vessel.
• Remove all debris, build-up, and detergent from the vessel before autoclaving.
Failure to do so may result in permanent etching or hazing of the vessel.
2. Attach a male Luer Fitting (r) to a length of Tygon Tubing (p).
3. Connect this tubing to the Culture Vessel (h) Female Luer fittings.
4. Cap or pinch-off the Tygon tubing (p) with a hose clamp (not included).
5. Place Stir Bar (j) in the Culture Vessel (h).
6. Add media to the Culture Vessel (h). Maximum volume should not exceed 600 ml.
7. Insert the Black O-Ring (l) into the Cap (o), and screw the Cap (o) onto the Culture
Vessel (h).
8. Insert the Red O-Ring (n) into top of the Cap (o), and place the Glass Lens (m) on top of
the Red O-Ring (n).
9. The Culture Vessel Assembly can now be autoclaved. The vessel may also be sterilized
using alternate methods such as rinsing with alcohol, dilute bleach solution, UV
irradiation, etc. The suitability of the sterilization method must be determined by the
user.
DO NOT AUTOCLAVE THE VESSEL WITH THE LED ASSEMBLY ATTACHED TO THE CAP.
DOING SO WILL DESTROY THE LED ASSEMBLY AND VOID THE WARRANTY.
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10. After the Culture Vessel Assembly has been autoclaved and allowed to cool, place the
Culture Vessel Assembly inside the Jacket Assembly (b) as shown in Figure 12 and
ensure a snug fit by pushing down with light force. Notice that the Vessel Ports point
toward you and the raised surfaces on either side of the vessel fit in the spaces
between the Jacket Assembly (b) halves.
11. Place the LED Assembly (c) on top of the Glass Lens (m) and turn it to engage its
locking pins into the mating lugs in the Cap (o), as shown in Figure 10.
12. Once the Vessel is inserted into the Jacket Assembly (b), do to remove the Jacket
Assembly from the PBR101 without first removing the Culture Vessel (h). Doing so
may result in damage to the wired components attached to the Culture Vessel
(probes, tubing, etc.).
Figure 9: Order of assembly of the Culture Vessel and Cap components
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PBR101 Photo Bioreactor
Guide to Setup and Operations
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Figure 10: Proper engagement of the LED Assembly locking pins onto Cap
Customizing the Culture Vessel Port Configuration
An advantage of the polycarbonate vessel is that the port configuration can be easily
customized by the user based on his/her specific requirements. This is not possible with a
glass vessel. In addition, the cost of the vessel is a small fraction of the cost of a glass
vessel. Because of this, most users find it convenient to have extra vessels on hand and
configured for specific experiments. For example, the standard vessel with 1-luer port in
the vessel for gas input, an extra vessel fitted with additional luer ports for use with the
Turbidistat Pump, and even variations of either of these (sampling ports). Because the
culture volume is also infinitely variable (between approximately 150 – 600 ml), additional
vessels with ports at different heights can also be of benefit.
Vessels may be custom configured when ordering, or they can easily be configured by the
user. A luer fitting may be added to the Vessel anywhere on the side of the Vessel that fits
between the Jacket Assembly halves, either on the front or back of the Vessel. The only
restriction is that the fitting cannot interfere with the OD sensors or cap threads.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
To add a luer fitting, follow these instructions:
1. Carefully drill a 5/32” hole using a sharp bit. Make certain the hole is drilled
perpendicular to the side surface of the vessel.
2. Remove all debris from the hole and surrounding surface.
3. Carefully tap the hole using a 10-32” tap. Make certain the tap is perpendicular to
the side surface of the vessel.
4. Add a small amount of solvent weld plastic bonding adhesive to the threads of the
fitting. Make certain adhesive does not get into the fitting passage way.
5. Thread the fitting into the threaded hole until it is fully seated. Finger tighten only!
Do not use a wrench or excessive force to tighten the fitting as this may result in
damage to the vessel and the fitting. The solvent adhesive will ensure a permanent,
leak-proof seal.
6. Let the solvent fully cure prior to use.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
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Figure 11: Vessel Assembly showing standard 1-port configuration and 4-port
configuration for use with the TP2501 Turbidistat Pump
Standard 1-port configuration 4-port configuration for use with the
TP2501 Turbidistat Pump
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PBR101 Photo Bioreactor
Guide to Setup and Operations
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Figure 12: Proper placement and orientation of the Vessel
Assembly within the Jacket Assembly (b)
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PBR101 Photo Bioreactor
Guide to Setup and Operations
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4.5 Attaching pH and Reference Electrodes, and Culture Temperature Probe
pH is monitored by using two half-cells. The half-cells consist of a sensor electrode and
reference electrode.
The pH Electrode (t) and Reference Electrode (u), both optional accessories, are distinguished
from each other by observing the tip design of each probe (see Figure 13).
Notice the Reference Electrode (right side) has a hole in the tip and white ceramic frit that
extends about 5-10 mm into the glass tip. The frit may vary slightly in color (white, off-white, or
beige) and length; this is normal and does not affect its performance.
The tip of the pH Electrode consists of a solid, translucent glass bulb which may be green to
amber in color.
Figure 13: Distinguishing between the pH (Left) and Reference Electrode (Right)
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Guide to Setup and Operations
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Prior to shipment, the pH and Reference Electrodes were conditioned and calibrated at the
factory to ensure proper performance. However, during shipping the probes may become
dehydrated and must be reconditioned prior to first use. Failure to do so may result in failure
of the probes to calibrate properly (see the Algal Command Software Manual for probe
calibration procedure) and/or perform properly during use. Reconditioning may also be
necessary periodically between uses in order to maintain performance. Before installing the
probes, please recondition them using the following simple procedure.
1. pH Electrode (t):
(a) Soak the pH Electrode (t) in 0.1N HCl for 20 minutes.
(b) Rinse in deionized or distilled water.
(c) The electrode is now ready for calibration.
2. Reference Electrode (u)
(a) Soak the Reference Electrode (u) in 1M KCl solution at 60-80°C for 20 minutes.
(b) Rinse in deionized or distilled water.
(c) The electrode is now ready for calibration.
3. Place both electrodes into pH 7 standard pH buffer solution (user supplied) and
immediately calibrate using the calibration routine in Algal Command (see the Algal
Command Software Manual included on this CD).
After successfully calibrating the electrodes, they may be installed in the Culture Vessel (h)
as follows:
1. Remove the stainless steel socket cap screws from the Cap (o) of the Culture Vessel (h).
Do not discard the cap screws. They may be replaced in the cap to prevent
contamination when autoclaving or when not using the pH and Reference Electrodes
during an experiment.
2. Insert the pH and Reference Electrodes into the Cap.
3. Screw the Electrodes into the Cap and tighten by hand only, being careful to allow the
cord to freely rotate without twisting or kinking. Failure to do so may result in
damage to the probe cable. If the threading is tight, use a small amount of water or
glycerin to provide lubrication.
4. Connect the pH and Reference Electrode cables to their corresponding BNC connectors
on the Control Tower.
5. Insert the Culture Temperature Probe (v) into the Cap in the remaining threaded hole.
6. Screw the Culture Temperature Probe (v) into the Cap and tighten by hand, being
careful to allow the cord to freely rotate without twisting or kinking. Failure to do so
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may result in damage to the cable.
7. Connect the Culture Temperature Probe (v) cable to the port labeled “Culture Temp” on
the Control Tower.
pH and Reference Probe Care and Maintenance
With proper probe care and maintenance the pH and Reference Electrodes will perform
well over time, maintaining responsiveness with minimal drift. Please observe the following
guidelines:
1. Probes should never be allowed to dry out. When not in use the tips should be
immersed in an electrode storage buffer (not included).
2. The pH (t), Reference (u), and Culture Temperature Probe (v) are not autoclavable.
They must be removed prior to autoclaving and sterilized using an alternate
method (the socket cap screws may be reinstalled in the cap prior to autoclaving in
order to maintain sterility of the media).
3. For optimum performance, the pH and reference electrodes should be recalibrated
daily. If this is not possible and they are used continuously within a culture over an
extended period, drift is to be expected. This is normal behavior for any pH probe.
4. After any extended use, it is recommended that the pH and reference probes be
reconditioned and recalibrated prior to use in a new experiment.
5. After extended use in a biological culture, it is expected that performance will
degrade due to contamination of the pH bulb and reference junction (frit).
Reconditioning will typically return these to factory performance, although this is
not certain and is a function of the user’s application. Reconditioning several times,
or using more rigorous procedures may be necessary after extended use or use in
biologically or chemically harsh conditions.
6. Exposure to conditions that promote fouling of the reference junction (e.g.
microscopic buildup of protein, lipid, cellular or otherwise any biological debris, salt
precipitation, etc.) will cause degradation of performance (drift, lack of
responsiveness, etc.). However, fouling may be reversed by careful removal of these
contaminants using appropriate cleaning procedures (e.g. solvents, detergents, acid,
heat, etc.). The specific method will depend on the fouling agent and guided by
knowledge of the culture components and conditions of the experiment.
7. Probes are considered semi-consumable components. Their useful lifetime will
depend on the application, length of exposure to fouling environments, frequency
of reconditioning, and user diligence to proper care and maintenance.
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4.6 Connecting the LED Assembly Power Cables
The LED Assembly provides light to the culture. The overhead light faithfully mimics
environmental sunlight interaction with an outdoor raceway pond. Using a shallow column
height, the light may also mimic light interaction with an enclosed tube bioreactor.
The LED Assembly is capable of providing light intensity greater than 3,000 microEinsteins at
the surface of the culture. Light intensity can be programmed using Algal Command to simulate
a diurnal cycle with varying lengths of day/night, intensity, etc.
• Plug the power cord of the LED Assembly (c) into the “Light 1” port on the Control
Tower (a).
• Plug the auxiliary cord of the LED Assembly (c) into the “Light 2” port on the Control
Tower (a).
• Calibrate the LED using the solar calibration routine in Algal Command Software and a
PAR meter (user supplied). See the Algal Command Software Manual for calibration
instructions. Solar Calibration must be performed prior to using the PBR101.
• Once the LED is calibrated, place the LED Assembly (c) on top of the Glass Lens (m) of
the Cap (o).
• Turn the LED Assembly (c) clockwise until it securely locks into the Cap (o) as shown in
Figure 10.
The LED Assembly must not be exposed to liquids. Doing so can damage the LED and cause
electric shock and fire. The LED assembly must be removed from the Vessel Cap prior to
autoclaving. There are no field serviceable parts in the LED Assembly. Removing the heat
sink or otherwise opening the LED Assembly may result in electric shock or fire and will void
the Warranty.
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4.7 Connecting Gas to the PBR101 and Culture Vessel
The PBR101 is supplied with a computer controlled gas solenoid valve. It will open and close to
deliver gas as instructed by Algal Command. See the Algal Command Program Manual for
instructions on programming the valve.
An optional additional valve may also be installed for supply of a second gas.
Inlet pressure to the PBR101 must not exceed 25 psi. Exceeding this limit may result in
damage and personal injury due to explosion of the tubing and valve.
In order to prevent excessive pressure, but while ensuring that each PBR101 has adequate gas
volume, it is recommended that gas be supplied by a high-pressure central source. Each
PBR101 should also be equipped (user supplied) by a second regulator limiting the PBR101
input pressure to 25 psi.
Attach the gas line from the low pressure regulator (user supplied) to the PBR101 as follows:
• Use tubing of appropriate pressure rating with an inside diameter of 1/8” to make a
connection between the regulator and the inlet fitting on the PBR101, as shown in
Figure 14. The inlet is on the left side of the Control Tower (a).
• Do not exceed 25 psi when connecting the gas source to the inlet fitting.
• Uncap/unpinch the Tygon tubing attached to the bottom of the Culture Vessel, and
attach this tubing to the outlet fitting on the PBR101, as shown in Figure 14. The outlet
is on the right side of the Control Tower (a). It is recommended that a 0.45 or 0.2
micron filter (user supplied) be placed in-line with this connection to prevent
contamination.
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Figure 14: Gas inlet (left panel) and outlet (right panel) connections to the PBR101.
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4.8 Powering On the PBR101
• Launch the Algal Command program on a computer (user supplied) connected to the
PBR101 via the supplied network Router (d). Minimum computer requirements are
provided in the Algal Command Software Manual.
• Ensure that the red voltage switch on the power supply is set to the correct line
voltage (115V or 230V). See Figure 3. Failure to set the voltage switch to the correct
line voltage will cause damage to the PBR101, electric shock, and fire.
• Turn the power switch of the power supply to the on position (represented by the |
symbol). See Figure 3.
• Within a few moments an icon will appear in the Algal Command Software that
represents the PBR101.
• See the Algal Command Software Manual for programming and controlling the PBR101.
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Guide to Setup and Operations
Rev. 0115
5. Setup and Operation of the TP2501 Turbidistat Pump
5.1 Key Features of the TP2501
• Allows for conducting turbidistatic experiments by simultaneously removing and
replacing exactly the same volume of media per unit time.
• Dilution can be triggered automatically by Algal Command based on a chosen target OD.
• Infinitely variable flow control from 0.875 – 8.910 ml/min permits infinitely variable
dilution rates.
• Single head, dual channel ensures in and out flows are perfectly matched when using
the same tubing for each channel (supplied).
• Simple plug and play – a single cable provides both power and programmable control
from the Algal Command Software Suite. No additional external power is required.
5.2 Parts Identification
If you ordered the optional Turbidistat Pump Module (cat. no. TP2501), you will receive the
additional components listed in Table 3 and shown in Figure 15.
Table 3: Turbidistat Pump components. Compare to Figure 15.
Item Quantity Note
w Turbidistat pump 1
x Pump Cable 1 Provides both power and
software control
y Luer Caps 4 Cap ports when not using the
pump
z Sampling Port 1 Septum for sterile inoculation
and/or sampling
aa Turbidistat Pump Tubing 2 Includes luer fittings
bb 4 Port Vessel 1 Ports for inflow of gas, sampling,
pump inflow, and pump outflow
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
Figure 15: Turbidistat pump components
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
5.3 Assembly
The following steps describe assembly of the TP2501 Turbidistat Pump:
1. Attach the Turbidistat Pump Tubing (aa) to the Culture Vessel (h) during the Culture
Vessel Assembly preparation, and cap or pinch the unattached end.
2. The typical setup with the-port vessel is show in the following picture:
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
3. Apply a small amount of silicone grease to the roller of the pump head.
4. After sterilization of the Culture Vessel Assembly, insert the Tubing (aa) onto the head
of the Turbidistat Pump (w) as described in the following steps:
(a) Push down on the bottom tab of the pump head, as shown:
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
(b) Slide both pieces of tubing onto the pump head as shown below. The spring clips
must be pulled out in order to seat the tubing onto the roller. One piece of tubing
will be in front of the other on the rollers.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
(c) Clamp each piece of tubing on both sides of the pump head using the two sets of
spring clips. Push the retaining tab (from step 1) up until it locks into position (may
hear it “click” into position). Once the tubing is loaded, it should appear as in the
following picture:
(d) Uncap the ends of the tubing.
(e) Place the tubing that will flow into the vessel into a container of sterile media.
(f) Place the tubing that will flow out of the vessel into a container for waste media.
5. Connect the Pump Cable (x) to the jack located on the back of the Turbidistat Pump (w).
6. Set the directional control switch on the Turbidistat Pump to the middle position. See
Section 5.4.
7. Install Algal Command Software before connecting the pump to the PBR101.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
8. Connect the other end of the Pump Cable (x) to the “Aux Out” port of the Control
Tower.
9. In Algal Command, set the “Aux Out” button to “On”. This will send power to the
turbidistat pump.
10. Set the directional switch to set the pump to remove liquid from the Culture Vessel, and
press the Purge button to prime the pump.
11. Once the Turbidistat Pump is primed and all air is purged from the lines, click the “Aux
Out” button in Algal Command so that the button reads “Off”. This will stop the pump
head from turning.
(a) Optionally set the directional switch on the Turbidistat Pump to the middle position
to stop the pump head.
12. Please see the Algal Command Software User Guide for instructions for Turbidistat
Pump programming and use during an experiment.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
5.4 TP2501 On-Board Operational Controls
1. Tubing retaining spring clips. Used to ensure tubing stays tight and in place against
roller head.
2. Pump head. Constructed of five durable polymer rollers to ensure smooth,
homogeneous fluid delivery. Rollers should be lubricated frequently using silicon grease
in order to prolong tubing life.
3. Retaining clip. Push down to load tubing; push up to lock tubing onto pump head.
4. Power indicator light. Illuminates when pump is running.
5. Purge button. Pushing this causes the pump head to rotate at maximum speed in
order to permit rapid purging of the lines. Purging can also be activated within Algal
Command.
6. Variable speed control. Allows for infinitely variable user controlled speed. This
permits users to finely tune the rate of culture dilution for a given experiment.
Frequency of pump triggering is programmed within Algal command based on the user
defined optical density.
7. Directional control switch. This switch allows the pump head to turn in either
direction. The middle position turns off head rotation.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
6. Maintenance Procedures
6.1 Cleaning External Surfaces
The PBR101 is finished with a very durable powder-coat epoxy enamel that is impervious to
most solvents, acids, bases, etc. Use only a soft damp cloth or paper towel for surface cleaning.
However, any liquid spilled on the PBR101 must be removed immediately to prevent damage
to internal components. Failure to do so can lead to electric shock and permanent equipment
failure.
6.2 Temperature Jacket Assembly
The insulation on the jacket assembly is important for maintaining accurate temperature
control of the vessel. Periodically inspect the insulation on the aluminum jacket assembly. Use
a very small amount of cyanoacrylate (CA) glue to repair any insulation that is peeling or
damaged. Caution: care must be exercised when using CA glue. Please follow manufacturer’s
instructions and precautions.
Periodically check wiring and connectors to ensure they are in good condition.
6.3 Tubing
Periodically inspect all tubing for cracks, bulges, and other forms of normal wear. Replace as
needed.
6.4 TP2501 Turbidistat Pump
Periodically grease the rollers with silicone roller grease formulated for this purpose. Also
periodically inspection and replacement worn tubing. It is recommended that tubing NOT be
draped over any part of the PBR101 or TP2501 where a leak could cause damage to electrical
components.
6.5 pH and Reference Electrodes (also see Section 4.5)
It is important to condition the electrodes prior to use and calibration of the PBR101. The
electrodes were conditioned and tested at the factor to verify their functionality. However,
during shipping and storage, the electrodes may dry out and require reconditioning.
Conditioning is simple and performed as follows:
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
pH electrode:
Place the bulb in 0.1 N HCl for about 20 minutes. Clean any debris by very gently wiping
the bulb with a Kimwipe. Take care not to scratch the glass bulb. Rinse with DI water prior
to calibration.
Reference electrode:
Place the tip in 1 M KCl solution at 60-80°C for 20 min. Rinse with DI water prior to
calibration.
The electrodes are sealed and do not require addition of electrode buffer. Maintaining the
electrodes by periodic reconditioning will greatly extend their useful life. Proteins, lipids,
minerals, salts, all organic matter present in culture will eventually lead to degradation of
electrode performance. Periodic reconditioning is important to maintaining good performance
and minimizing drift, slow response, etc. For best performance, daily recalibration and/or
conditioning (depending on the use) is recommended.
Periodically check electrode cable insulation and threaded fittings for normal wear. Repair or
replace as necessary.
The pH and Reference Electrodes are NOT autoclavable. Autoclaving may damage the cable.
Electrodes may be sterilized using ethanol, mild bleach, isopropanol, and other commercially
available sterilizing solutions.
When not in use, electrodes should be stored in commonly available electrode storage buffer
(not supplied). Alternatively, the pH and Reference Electrodes may be stored in pH 4.01
standard buffer with 1/100 part of saturated KCl added. Do not allow either electrode to dry
out completely. If storing for an extended period, exchange the storage buffer every 3 months.
Never store electrodes in water or other weakly buffered solutions.
6.6 Culture Temperature Probe
Periodically inspect and clean any debris or scale from the probe. This is recommended after
each experiment. The probe is made from stainless steel and resistant to damage from culture
components, but scale and debris removal will ensure optimum performance.
The Culture Temperature Probe is NOT autoclavable. Autoclaving may damage the cable.
The Culture Temperature Probe may be sterilized using ethanol, mild bleach, isopropanol, and
other commercially available sterilizing solutions.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
6.7 Culture Vessel
The Culture Vessel is made from durable, clear polycarbonate (PC) and can be cleaned with
laboratory detergents that are safe for use on PC. Use only a soft sponge or bottle brush and
hot water to avoid scratching the vessel. Scratching can lead to premature failure and cracking,
and effect OD readings which are made through the vessel wall. Use only LOW heat or air
drying since high heat can cause warping and failure.
The Vessel may be autoclaved several times. However, after several cleaning and autoclaving
cycles, the vessel may become slightly hazy or cloudy. This is considered normal wear. The
vessel is considered a semi-consumable component and should be replaced once hazing occurs
or the vessel otherwise shows signs of wear (cracks, scratches, etc.). Replacement vessels are
low cost, readily available (catalog number CUVL102), and customizable based on user
requirements (see below). With proper care, the vessel should last for a considerable number
of experiments.
Polycarbonate is also is resistant to a variety of commonly used sterilizing agents including
ethanol, isopropanol, mild bleach solution, and most commercially available sterilizing
solutions.
It is important to wash debris from the vessel immediately upon completing an experiment.
Debris left to dry can become difficult to remove and may ruin the vessel. Further, autoclaving
a dirty vessel on a dry cycle may permanently bond the debris to vessel and/or etch the vessel
wall.
Periodically inspect the vessel for cracks, loose or worn fittings. Replace if required.
6.8 Electrical Cables
Periodically inspect all electrical cables for fraying, loose connections, and worn insulation.
Replace as needed.
6.10 Cap Assembly
The Cap and other components of the Vessel Assembly (O-rings and glass lens) can be cleaned
and sterilized using the same treatments used for the Vessel.
Periodically clean the Glass Lens in order to ensure maximum light transmission into the vessel.
Use any mild laboratory glassware detergent and a soft sponge or brush. Do not scratch!
Periodically inspect the O-rings, glass window, and cap for wear. The integrity of these
components is critical to avoiding culture contamination. Replace as needed.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
7. Consumables and Field Serviceable Parts
The follow is a list of the most commonly required field serviceable replacement and
consumable parts for the PBR101. Many users find it convenient to keep spares on-hand.
Part (see Figures 1 and 2 for identification) Catalog number
Culture Vessel (h)* CUVL102
Culture Vessel cap (o) CVLC102
Female Luer fittings (s) LUF102
Male Luer fittings (r) LUBM102
Luer Caps (q) LUCP102
Gas inlet/outlet tubing (p)
Black O-ring (l) BOR102
Red O-ring (n) ROR102
Glass Lens (m) VLWW102
Stir bar (j) STBR102
Reference electrode (u) PHRP102
pH Electrode (t) PHSP102
Preventative Maintenance Kit: includes black and red O-
rings, glass lens, luer fittings and caps. PMK102
Turbidistat Pump Tubing TPTB102
*Vessels may be ordered with our standard port configuration or with virtually
any customer-specified configuration.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
8. Trouble-Shooting and Technical Support
Phenometrics’ technical support staff is expertly trained in the engineering, setup, operation,
maintenance, repair, and use of Phenometrics’ instruments. Should you have any questions or
issues about the setup, maintenance, or operation of your PBR101, please contact technical
support at:
+1-517-884-4361
The following table lists some of the most common issues and solutions fielded by our support
department.
Unit will not power on Power not connected Check power source and that power
switch is turned to ON
Voltage switch not set properly Check that voltage switch is set to
proper line voltage
Ethernet cable not connected Ensure that the Ethernet cable is
securely connected and that PBR is
recognized by algal command (see
the Algal Command Software manual)
Culture contamination during
experiment
Incorrect vessel assembly or
worn parts
Inspect all cap components and
ensure an airtight seal when LED
assembly is locked in place.
Ensure O-rings are properly seated
and not worn
Ensure an airtight seal of all gas
connections. Filter BOTH in and out
gas streams (0.2 micro inline filter
recommended).
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
Poor sampling or inoculating
technique
Validate aseptic technique when
preparing media, inoculating,
sampling. Septum, sterile needle and
syringe, sterilize septum prior to
piercing with needle, etc.
Ensure inoculum is not contaminated.
Verify independently using
appropriate microbiology technique
such plating, etc.
Poor sterilization Validate sterilization procedure and
equipment, including autoclave. Use
autoclave test tape or capsules to
ensure complete sterilization.
Autoclave vessel assembly with
media on liquid cycle. Ensure
temperature probe and pH
electrodes are properly sterilized and
use good technique when inserting
them into sterilized vessel.
Vessel temperature not being
maintained
Jack temperature probe Verify installation
Verify probe function with a VOM.
The thermistor should give a
resistance of approximately 10K-Ohm
at 25°C.
Replace probe
Vessel temperature probe Verify installation
Verify probe function with a VOM.
The thermistor should give a
resistance of approximately 10K-Ohm
at 25°C.
Jacket assembly Verify that the jacket assembly is
connected to the control tower.
Make sure that fans are being
powered, and are moving air.
Make sure that all electrical
connections are clean and free of
corrosion.
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PBR101 Photo Bioreactor
Guide to Setup and Operations
Rev. 0115
On-board fuse has tripped Remove anything plugged into the
Aux Out port. Turn off PBR101 and
reboot to automatically reset the
fuse.
If problem persists, contact Technical
Support.
pH probe will not give linear
calibration, slow response, or
shows significant drift
Contaminated/dirty reference
and/or pH sensor
Recondition electrodes and
recalibrate (see Maintenance
section).
Electrodes permanently fouled
or at end of life
Replace reference and/or pH
electrode
Application Some applications necessitate
frequent reconditioning and/or
recalibration. For best results, daily
recalibration is recommended.
Control and response of LED
is lost
Improper connection Ensure connections according to
instructions
Lose of communication to Algal
Command
Ensure that PBR101 is connected to
the Algal Command Software.
Stir bar vibrates and/or does
not spin
Increasing speed too rapidly Turn speed to 0 and slowly increase
to desired speed in increments of 50
rpm
Motor may occasionally stick and
need slight manual rotation.
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