Photon Systems Instruments FMT150 Guide

Photobioreactor
FMT150
Manual and User’s Guide
Please read this manual before operating this product
PSI, spol. s r. o., Drásov 470, 664 24 Drásov, Czech Republic
FAX: [phone], TEL: [phone], www.psi.cz

© PSI (Photon Systems Instruments), spol. s r. o.
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©PSI (Photon Systems Instruments), spol. s r. o. (hereinafter PSI), 2019
This document and its parts can be copied or provided to a third party only with the express permission of PSI.
The contents of this manual have been verified to correspond to the specifications of the device. However, deviations cannot
be ruled out. Therefore, a complete correspondence between the manual and the real device cannot be guaranteed. The
information in this manual is regularly checked, and corrections may be made in subsequent versions.
The visualizations shown in this manual are only illustrative.
This manual is an integral part of the purchase and delivery of equipment and its accessories and both Parties must abide
by it.

© PSI (Photon Systems Instruments), spol. s r. o.
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TABLE OF CONTENTS
1Warnings and Safety Precautions ................................................................................................................................... 5
2General Description ........................................................................................................................................................ 7
3Device Description .......................................................................................................................................................... 8
3.1 Configurations and Versions.................................................................................................................................. 9
3.2 Standard Components......................................................................................................................................... 14
3.3 Control Unit Front Panel...................................................................................................................................... 16
3.4 Rear Panel For Version FMT150.1 ....................................................................................................................... 18
3.5 Rear Panel For Version FMT150.2 ....................................................................................................................... 20
3.6 Connection Schema............................................................................................................................................. 22
4Device Installation......................................................................................................................................................... 23
4.1 Power Supply....................................................................................................................................................... 23
4.2 Cultivation Vessel ................................................................................................................................................ 24
4.3 pH/Temperature Sensor...................................................................................................................................... 28
4.4 Gas Supply ........................................................................................................................................................... 31
5Optional Accessories/Components............................................................................................................................... 33
5.1 PBR Software....................................................................................................................................................... 33
5.2 Air Pump With Bubble Interruption Valve........................................................................................................... 33
5.3 pH/Temperature Module .................................................................................................................................... 34
5.4 O2Module ........................................................................................................................................................... 34
5.5 CO2Module ......................................................................................................................................................... 36
5.6 Magnetic Stirrer................................................................................................................................................... 43
5.7 PWM Pump.......................................................................................................................................................... 46
5.8 Gas Mixing System GMS 150 ............................................................................................................................... 48
5.9 Gas Analysis System GAS 150.............................................................................................................................. 51
5.10 Peristaltic Pump PP600........................................................................................................................................ 54
5.11 External Light Panel............................................................................................................................................. 58
5.12 Additional Software Licence................................................................................................................................ 59
5.13 Light Upgrade ...................................................................................................................................................... 59
5.14 Supplementary/Replacement Components........................................................................................................ 59

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6Cultivation Guide .......................................................................................................................................................... 60
6.1 Set-Up the Components ...................................................................................................................................... 60
6.2 Set-Up the Experiment ........................................................................................................................................ 60
7Appendix ....................................................................................................................................................................... 62
7.1 FMT150 Control Menu Tree ................................................................................................................................ 62
7.2 QY Measurement ................................................................................................................................................ 70
7.3 Technical Specification Standards ....................................................................................................................... 71
8Warranty Terms and Conditions ................................................................................................................................... 75
9Troubleshooting and Customer Support....................................................................................................................... 76
10 List of Abreviations .................................................................................................................................................. 77

© PSI (Photon Systems Instruments), spol. s r. o.
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1WARNINGS AND SAFETY PRECAUTIONS
PLEASE READ THE FOLLOWING INSTRUCTIONS CAREFULLY BEFORE TURNING THE PHOTOBIOREACTOR ON:
•Remove all packaging and transport protectors before connecting the Photobioreactor to the power supply.
•Use only cables supplied by the manufacturer.
•Keep the device dry outside and avoid working in high humidity environment!
•The manufacturer is not responsible for any damage due to improper operation!
•Water and other liquids should only be placed in vessels designed for the purpose and according to instructions
included in this manual.
GENERAL ELECTRICAL SAFETY GUIDELINES:
•Perform a routine check of the devices and their wiring.
•Replace worn or damaged cords immediately.
•Use appropriate electrical extension cords/power bars and do not overload them.
•Place the device on a flat and firm surface. Keep away from wet floors and counters.
•Avoid touching the device, socket outlets or switches if your hands are wet.
•Do not perform any alterations to the electrical parts of the device or its components.
The following table presents basic highlight symbols used in this manual:
Symbol
Description
Important information, read carefully.
Complementary and additional information.
Tab. 1 Used symbols.

© PSI (Photon Systems Instruments), spol. s r. o.
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The Photobioreactor is considered Class 1M* LED Product. LED radiation may be harmful to eye.
Avoid direct and strongly reflected exposure. Use protective glasses.
*Class 1M: Laser and LED equipment that is safe for the naked eye under foreseeable conditions of operation.
Looking directly into the source of radiation by employing optics within the beam, such as magnifying glass,
telescope or microscope, can be potentially hazardous.

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2GENERAL DESCRIPTION
Photobioreactor FMT150 feature a unique combination of the cultivator and computer controlled monitoring device. The
Photobioreactor FMT150 combines a cultivation vessel with the built-in fluorometer and densitometer. It is primarily
designed for modular and precise phototrophic cultivation of algae, cyanobacteria and bacteria. Light power and spectral
composition as well as temperature and aeration gas composition can be set with high accuracy.
User defined cultivation conditions can be dynamically varied according to user defined protocol. Continuous online
monitoring of cultivation conditions and measured parameters is possible via the optional PBR software. Light,
temperature, gas composition, and culture medium regime can be programmed to oscillate with various amplitudes and
frequencies with user-defined time steps from seconds to hours via the software or via the control unit. Turbidostatic
cultivation can be used for the stabilization of the suspensions via optical density control. In addition, up to 8 peristaltic
pumps for different chemostat cultivation control can be linked to the Photobioreactor FMT150 for the highly precise
control of cultivation conditions.
The growth of the cultures is monitored by the integrated densitometer (OD 720, 680 nm). Relative chlorophyll content of
the culture can also be monitored continuously by the difference of optical densities at 680 and 720 nm. The instantaneous
physiological state of the culture is measured by the Photosystem II quantum yield Fv/Fm.
The Photobioreactor FMT150 has a flat-vessel design that enables uniform illumination over the whole culture.
Photobioreactors are currently manufactured in three cultivation vessel capacities: 400 ml, 1000 ml and 3000 ml.
All Photobioreactor models may be supplied with a number of optional accessories –sensors, stirrers, pumps, etc. Control
of the Photobioreactor is possible with either the control unit of the photobioreactor itself or the PBR software. However,
the PBR software provides much wider functionality to define, control, display and analyze the experimental conditions
and data.

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3DEVICE DESCRIPTION
Standard version of the Photobioreactor FMT150 consists of a cultivation vessel with a sealable lid and a base box
containing electronics circuitry, LED light panel (Fig. 1A) and other components essential for the optimal operation of the
photobioreactor.
Fig. 1 A) Photobioreactor body consists of a flat vessel holding the suspension and of a base box with electronic circuitry. B) The block
scheme of the instrument shows the Control Processor Unit exchanging data and protocol with external computer (PC) and auxiliary
external modules. The Control Unit governs measurements of parameters such as OD and chlorophyll fluorescence plus parameters
measured by auxiliary sensors. It also controls the actinic light sources and temperature of the culture. C) Detail of the LED
configuration. D) Uncorrected optical density spectrum of cyanobacterial suspension. The lines with arrows indicate wavelengths of
various light sources that are integrated in the instrument. The detection range of the photodiode is also shown. E) Detail (rotated
top-to-bottom view) of the fluorometer and densitometer. F) Schematic representation of optional gas supply components and
modules that can be used to stabilize aeration and set CO2concentration level.

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The array of high-power light emitting diodes (LEDs) is located behind the cultivation vessel.
In the bottom corner of the front of the vessel is a semiconductor light sensor for measuring fluorescence emission and
suspension optical density by attenuation of light that was emitted from the LEDs (Fig. 1E)
The Photobioreactor can also control external modules such as peristaltic pumps and solenoid valves via the
photobioreactor control unit (shown schematically in Fig. 1F and Fig. 2). The solenoid valves are used to switch off the gas
supply to the culture (bubbling) during optical measurements. Peristaltic pumps are used in turbidostat and chemostat
modes to supply fresh medium or buffer.
Fig. 2 Schematic diagram of the Photobioreactor, its modules and remote control.
3.1 CONFIGURATIONS AND VERSIONS
FMT150 can be shipped with various configurations of vessel capacity, light quality, light intensity and thermoregulation.
Numerous additional accessories such as various sensors can be supplied as described in chapter 5 on page 33.
Please note that this manual describes operation of two FMT150 versions (Fig. 4) - older PBR FMT150.1 and new PBR
FMT150.2.
3.1.1 CULTIVATION VESSEL
The cultivation vessel is flat and rectangular in shape. Its front and back windows are made of glass plates. The base is
made out of stainless steel and contains a thermal bridge that facilitates heat transfer between a Peltier cell in the
instrument base and the culture suspension. The lid of the vessel is easily detachable during cleaning and contains Luer
connectors for gas or medium supply tubing, waste medium exhaust or sample collection. The three larger ports
accommodate the optional temperature/pH sensor and dissolved O2or CO2 probes.
The Photobioreactor FMT150 is manufactured in three vessel capacities enabling the customer to select optimal cultivation
conditions for his experiments.

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•400 ml (in FMT150/400)
•1000 ml (in FMT150/1000)
•3000 ml (in FMT150/3000)
3.1.2 LED LIGHTING
The high-power light emitting diodes generate a highly uniform irradiance flux that can be controlled in the range
0 –3,000 mol.m-2.s-1 PAR. The irradiance can be dynamically modulated by the FMT150 control unit or by the PBR
software. Spectra are shown in Fig. 3.
Two standard LED panel versions are available, each color is controlled independently:
•White-Red panel: maximum intensity 1,500 mol.m-2.s-1 (white 750 / red 750 mol.m-2.s-1)
•Blue-Red panel: maximum intensity 1,500 mol.m-2.s-1 (blue 750 / red 750 mol.m-2.s-1)
Two optional upgraded LED panel versions are available, each color is controlled independently:
•White-Red panel: maximum intensity 3,000 mol.m-2.s-1 (white 1,500 / red 1,500 mol.m-2.s-1)
•Blue-Red panel: maximum intensity 3,000 mol.m-2.s-1 (blue 1,500 / red 1,500 mol.m-2.s-1)
Optional customized panel versions:
•White-Blue: maximum intensity 1,500 mol.m-2.s-1
•White-Blue with Light Upgrade: maximum intensity 3,000 mol.m-2.s-1
•Red: maximum intensity 750 mol.m-2.s-1 (no light upgrade available)
•White: maximum intensity 1,500 mol.m-2.s-1
•White with Light Upgrade: maximum intensity 3,000 mol.m-2.s-1
•Blue: maximum intensity 1,500 mol.m-2.s-1
•Blue with Light Upgrade: maximum intensity 3,000 mol.m-2.s-1

© PSI (Photon Systems Instruments), spol. s r. o.
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Fig. 3 LED spectra of PBR FMT150.
3.1.3 THERMOREGULATION
It requires presence of the pH/Temperature probe.
•Standard range (ambient temperature around 23 °C):
5 –75 °C (FMT150/400)
10 –75 °C (FMT150/1000 and FMT150/3000)
Please find approximate estimation of time required for the given temperature equilibration in the Tab. 2 below.
Initial
temperature
Target
temperature
Light conditions
µmol.m-2.s-1
Equilibration time
required
5 °C
50 °C
1,500
2.5 hours
50 °C
5 °C
1,500
3 hours
Tab. 2 Temperature equilibration times.
3.1.4 VERSIONS
The available vessel capacity for two FMT150 versions:
•FMT150.1 –400 ml, 1000 ml
•FMT150.2 –400 ml, 1000 ml, 3000 ml
Important: The FMT150 in both 400 and 1000 ml models have been upgraded from FMT150.1 (Fig.
4A) to FMT150.2 (Fig. 4B). This manual provides instructions for operation of both versions (original,
now referred to FMT150.1 and the current version FMT150.2). Please always ensure that you refer to
correct instructions.

© PSI (Photon Systems Instruments), spol. s r. o.
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Fig. 4 Two versions of the PBR FMT150: A) FMT150.1, B) FMT150.2

© PSI (Photon Systems Instruments), spol. s r. o.
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List of parameters measured and/or calculated by the Photobioreactor FMT150:
Symbol
Name
Description
Ft (Dark)
Instantaneous fluorescence in the
dark
Instantaneous F-yield in dark equals F0
Ft = F0 (in dark-adapted sample)
Ft (Light)
Instantaneous fluorescence in the
light
Instantaneous F-yield in light that results from a dynamic
equilibrium of plastoquinone reducing and re-oxidizing
processes and from non-photochemical quenching
Ft = Fs (in light-adapted sample)
FM (Dark)
Maximum fluorescence in dark-
adapted state
Maximum F-yield with QAreduced (qP=0) and non-
photochemical quenching relaxed (NPQ=0)
FM (Light)
Maximum fluorescence in light-
adapted state
Maximum F-yield with QAreduced (qP=0) and non-
photochemical quenching active (NPQ≠0)
FV
Maximum variable fluorescence
FV = FM - F0
QY (Dark)
Instantaneous PSII quantum yield
in the dark
Effective PSII quantum yield
QY= FV / FM(dark-adapted sample)
QY (Light)
Instantaneous PSII quantum yield
in the light
Effective PSII quantum yield relaxing in light
QY= FV’ / FM’ (light-adapted sample)
OD680
Optical density at 680 nm
Optical density representing light scattering and chlorophyll
absorption.
OD720
Optical density at 720 nm
Optical density representing light scattering.
pH
pH
pH is the negative logarithm of the hydrogen ion
concentration in the medium.
[O2]
Oxygen concentration
Concentration of dissolved oxygen.
[CO2]
Carbon dioxide concentration
Concentration of dissolved carbon dioxide. CO2partial
pressure is calculated using the measured pH and the
temperature.
Tab. 3 FMT150 measured parameters.

© PSI (Photon Systems Instruments), spol. s r. o.
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3.2 STANDARD COMPONENTS
Photobioreactor FMT150 can be purchased in different configurations from minimal up to complex system consisting of
numerous accessories linked together. They can be controlled via the Photobioreactor control unit itself or by the PBR
software for full operation of the system. However, the Photobioreactor control unit offers only static manual control.
Minimal required configuration that is recommended to be purchased for optimal operation of the Photobioreactor
cultivation and monitoring system consists of set of standard photobioreactor components (as listed below) plus air pump
with bubble interruption valve and pH/temperature module. Components that are part of minimal Photobioreactor
FMT150 configuration are shown in Fig. 6. Optional components that are available as accessory modules and can be
purchased together with Photobioreactor FMT150 are described in chapter 5.
Photobioreactor FMT150 purchased in minimal configuration can be effectively used for cultivation and monitoring
purposes, however without purchasing PBR software no measured data can be stored and visualized as well as no user-
defined protocols can be designed. Chapter 3.3 and chapter 7.1 describe how to operate the PBR FMT150 in cultivation
mode via front panel control unit, without use of PBR software.
Note: Customers are kindly asked to check the contents of the delivered package and compare it with enclosed package
list generated based on customer‘s order. Please contact the su[email protected] if any discrepancies occur.
As shown on Fig. 6 on page 16 the FMT150 is shipped with:
1. Power supply with the power cord.
2. Photobioreactor with integrated double-modulation fluorometer- Ft, Fm, QY and integrated optical density
measurement.
3. Cultivation vessel with capacity 400 ml (with holder) for FMT150/400, 1000 ml for FMT150/1000 or 3000 ml for
FMT150/3000.
4. LED panel shield.
5. Spare Part Kit* –for the complete list of Spare Part Kit see Tab. 4 on page 15 and corresponding Fig. 5.
6. User guide (this document) and Software manual.
7. Bubble Humidifier –To maintain stable culture volume.
Components #8 and #9 below are highly recommended for proper functionality of the PBR, but are not included in the
standard configuration and have to be purchased separately.
8. Bubble interruption valve with air pump (air pump not shown on Fig. 6).
9. pH/temperature module.

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Fig. 5 Components of the Spare part kit. Numbers correspond to component number in Tab. 4.
Component
Number
Component Description
Specification
Explanation
Pcs
1
Male Luer Integral Lock MTLL240-
J1A*
5/32” (4.0 mm)
10
2
Female Luer Thread Style BSFTLL-
J1A*
5/16” Hex to 1/4-28
UNF
Bottom Sealing Thread
For fixing of stainless tubes in the
cuvette’s lid. The fittings are
modified - drilled with 3 mm drill bit
10
3
Female Luer Thread Style FTLL240-1*
5/32” (4.0 mm)
10
4
Male Luer Integral Lock Ring Plug
LP4-J1A*
10
5
Tee Tube Fitting T40-J1A*
5/32” (4.0 mm)
10
6
Reduction Tube Fitting 4060-1*
1/4” (6.4 mm) and
5/32” (4.0 mm)
5
7
Double Check Valve DCV118-001*
Safety valves that permit gases and
liquids to flow in only one direction
5
8
Check Valve SCV08053*
Check valve for with low priming
volume
5
9
Metal lid with ports (9a) with silicone
ring (9b)
Silicon ring 27x42x2
mm
For humidifier bottle
1+2
10
O-ring
75x3 mm
Sealing between lid and the vessel
5
11
O-ring
14x2 mm
Sealing to the openings for optional
sensors
10
12
Silicon Tubing
Ø 6/3 mm
Additional tubing
10 m
13
Silicone tubing
Ø 5/2 mm
Additional tubing
0.5 m
14
Hex Key
5 mm
To loosen up the screws of the lid
1
15
Spanner/Screwdriver
8 mm
For releasing filler screws in optional
sensor positions and luer connectors
in the vessel lid
1
16
T-tube (silicone tubing with 2 pcs of
Male Luer Lock and 1 pc of Tee Tube
Fitting)
Aeration tubing for the vessel
1
17
Wrench
19 mm
For releasing sensors
1
Tab. 4 List of Spare part kit –components shown in Fig. 5.
* Provided by: www.valueplastics.com

© PSI (Photon Systems Instruments), spol. s r. o.
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Fig. 6 Minimal configuration set of the Photobioreactor FMT150. Numbers correspond to component numbers as described on page 14.
3.3 CONTROL UNIT FRONT PANEL
Photobioreactor Control Unit front panel can be used for simple manual control of the PBR FMT150. By using the keys on
the front panel, the operation of cultivation and monitoring units such as pH/temperature sensor, light control, OD
monitoring module, PWM mode or CO2and O2sensor can be controlled. When FMT150 photobioreactor is purchased
without the Remote Control System for PBR FMT150 then all of the control has to be set up manually using the front panel
of the FMT150 control unit. The user can read out from the display the actual values for the given sensor or monitoring
unit.
Important: Please note that no user-defined protocols can be designed and no data recording is
possible without the PC and PBR software.
Please note that neither turbidostat nor chemostat module can be operated if FMT150 is purchased
without the PBR software.
The front panel of the FMT150 control unit has 4 LED lights on the left and 5 control keys on the right (Fig. 7).

© PSI (Photon Systems Instruments), spol. s r. o.
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Fig. 7 Photobioreactor front panel.
3.3.1 LED INDICATORS
Green LED indicator READY is lighting when the current temperature is equal to target temperature.
Orange LED indicator HEAT is lighting when the heater turns on in the cultivation vessel.
Blue LED indicator COOL is lighting when the water in the cultivation vessel is cooled and the Cooling Unit is operating. The
minimal temperature in the standard PBR FMT150 is 5 °C.
Red LED indicator OVERLOAD is lighting when the requested temperature in the cultivation vessel higher than the current
temperature was not reached yet.
3.3.2 DIGITAL DISPLAY ON THE LEFT
Displays the current temperature in the cultivation vessel and error messages if any problems with the FMT150 occur.
Possible error messages are:
•Er6 –Temperature/pH sensor is not connected.
•Er7 –Temperature sensor is not functional and must be repaired. Please contact su[email protected] if this error
occurs.
3.3.3 DISPLAY ON THE RIGHT
This display shows various functions of the FMT150 and these are activated and set with the main keys on the right side of
the control unit as described below.
3.3.4 MAIN KEYS
•[M]: Used to move back in the menu tree or to exit the menu.
•[S]: Used to move forward in the menu tree, to save the selection, or to turn ON/OFF.
•[▲]: Used to move up in the menu or to add value.
•[▼]: Used to move down in the menu or to subtract value.
•[Mains]: (On/Off) key doesn't have any function related to the common use of PBR and it is intended only for
service purposes. The Photobioreactor is turned on by switching on the corresponding Power supply.

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Please refer to the chapter 7.1 if Photobioreactor FMT150 is purchased without the PBR software and
the PBR FMT150 is to be controlled solely manually via the control unit front panel.
3.4 REAR PANEL FOR VERSION FMT150.1
The rear panel houses connectors for all connecting cables (Fig. 8).
Fig. 8 Photobioreactor control unit and its power supply rear panels. A) Back panel of the Photobioreactor control unit with the serial
number (blue arrow). Numbers correspond to the description on below. B) Rear panel of the power supply with the serial number (blue
arrow). Note the same serial number. Each Photobioreactor has its own power supply and it cannot be interchanged with another
power supply.
3.4.1 CONNECTOR FOR PH/TEMPERATURE SENSOR
pH/temperature sensor should be plugged to this connector when in use. PBR FMT150 control unit recognizes when the
sensor is connected and will automatically control the operation of the sensor.
3.4.2 CONNECTOR FOR OPTIONAL SENSOR
O2 sensor should be plugged to this connector when in use. PBR FMT150 control unit recognizes when the sensor is
connected and will automatically control the operation of the sensor.
Only O2 sensor can be plugged to this connector.

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3.4.3 SERIAL COMMUNICATION CONNECTOR
USB connector with serial communication cable is provided as part of the PBR FMT150 package when purchased with the
PBR software to connect the PBR FMT150 with the computer. This connection has to be made for the communication of
the PBR FMT150 via the PBR software.
3.4.4 AUX1 CONNECTOR
AUX1 connector should be used for the CO2sensor, peristaltic pump, additional light or magnetic stirrer (please see page
22 for detail description of possible connection schemes).
3.4.5 AUX2 CONNECTOR
AUX2 connector is used primarily for the bubble interruption valve, but can be used also for CO2sensor, peristaltic pump,
additional light or PWM pump (please see Fig. 10 page 22 for detail description of possible connection schemes).
3.4.6 POWER SUPPLY CONNECTOR
The power supply unit is plugged into the FMT150 control unit via this connector.
3.4.7 SERVICE COMMUNICATION CONNECTOR
Service communication connector is designed only for servicing the FMT150 by the manufacturer.
Warning: NEVER CONNECT THE SERVICE CABLE TO THIS CONNECTOR (RED ARROW IN Fig. 8A)
WITHOUT PRIOR CONSULTATION WITH THE MANUFACTURER.
3.4.8 IDENTIFICATION LABEL WITH SERIAL NUMBER
Each Photobioreactor FMT150 produced has a serial number that corresponds with the serial number of the power supply
unit. These are always provided together and are not interchangeable. Prior to switching PBR FMT150 device ON make
sure that the serial number on the rear panel of the main body matches the serial number on the power supply (Fig. 8B).
Warning: Damage can occur when the power supply is incompatible with the PBR FMT150 device.

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3.5 REAR PANEL FOR VERSION FMT150.2
The rear panel houses connectors for all connecting cables (Fig. 9). Please note that the numbers in the next section
correspond to the numbers in Fig. 9.
Fig. 9 Photobioreactor control unit and its power supply rear panel. A) Back panel of the Photobioreactor control unit. B) Serial number
on the front panel of the PBR FMT150 unit. C) Rear panel of the power supply with the serial number (blue arrow).
3.5.1 CONNECTOR FOR PH/TEMPERATURE SENSOR
pH/temperature sensor should be plugged to this connector when in use. PBR FMT150 control unit recognizes when the
sensor is connected and will automatically control the operation of the sensor.
3.5.2 CONNECTOR FOR O2SENSOR
O2sensor should be plugged to this connector when in use. PBR FMT150 control unit recognizes when the sensor is
connected and will automatically control the operation of the sensor.
3.5.3 USB PBR COMMUNICATION CONNECTOR
USB communication cable is provided as part of the PBR FMT150 package when purchased with the Remote Control System
to connect the PBR FMT150 with the computer. This connection has to be made for the communication of the PBR FMT150
via the PBR software.
3.5.4 AUX1 CONNECTOR
AUX1 connector should be used for the CO2sensor, peristaltic pump, PWM pump, additional light or magnetic stirrer
(please see Fig. 10 on page 22 for detail description of possible connection schemes).