PBR101 Photo Bioreactor Guide to Setup and OperationsPBR101 Photo Bioreactor Guide to Setup and...

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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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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• 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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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

[email protected]

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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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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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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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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• 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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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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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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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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Figure 4: Connection of Ethernet Cable (g) to Control Tower (a) and Router (d)

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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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• 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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• 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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• 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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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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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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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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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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Figure 12: Proper placement and orientation of the Vessel

Assembly within the Jacket Assembly (b)

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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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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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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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Figure 15: Turbidistat pump components

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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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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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(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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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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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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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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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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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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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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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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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:

[email protected]

+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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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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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.

44

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