Operating Manual
2019-02-07
Minifors 2
Benchtop Bioreactor
Dok-ID: 75623 / V.03.00 - Original
Infors AG
Headoffice, Schweiz
service@infors-ht.com
Infors GmbH
Infors UK Ltd
Infors Sarl
www.infors-ht.fr
Infors Benelux BV
infors.bnl@infors-ht.com
Infors USA Inc.
www.infors-ht.us
Infors Canada
Infors Bio-Technology
info@infors-ht.com.cn
Infors South East Asia
Infors LATAM Ltda.
Infors.br@infors-ht.com
www.infors-ht.com
Engineering and production in Switzerland
Rittergasse 27
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T +41 (0)61 425 77 00
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info@infors-ht.com
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The Courtyard Business Centre
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Velp (GLD)
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Wangjing New Industrial Zone
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Contact details of our local dealers worldwide can be found
on our website.
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Minifors 2 - Benchtop Bioreactor
Table of Contents
1 General Information ............................................................... 9
1.1 About this Manual .......................................................... 9
1.2 Explanation of Special Notices ...................................... 9
1.2.1 Warning Notices ............................................. 9
1.2.2 Other Notices................................................ 10
1.3 Equipment Identification (Standard Identification Plate)
..................................................................................... 10
1.4 Declaration of Conformity ............................................ 11
1.5 Customer Service and Services .................................. 11
2 Safety and Responsibility ................................................... 12
2.1 Intended Use, Incorrect Use and Misuse .................... 12
2.2 Qualified Personnel ..................................................... 13
2.2.1 Provider ........................................................ 13
2.2.2 User .............................................................. 13
2.2.3 Operator ....................................................... 14
2.3 Unauthorised Persons ................................................. 15
2.4 Responsibility of the Provider ...................................... 15
2.5 General Hazards .......................................................... 15
2.5.1 Electrical Current .......................................... 16
2.5.2 Unauthorised Spare Parts and Accessories 16
2.6 Particular Hazards ....................................................... 16
2.6.1 Hot Surfaces ................................................. 17
2.6.2 Dangerous Gases ........................................ 17
2.6.3 Flammable or Explosive Substances ........... 17
2.6.4 Corrosive or Toxic Substances .................... 17
2.6.5 Bioactive Substances or Pathogenic
Organisms .................................................... 18
2.6.6 Overpressure or Vacuum ............................. 18
2.7 Warning Symbols on the Equipment ........................... 18
2.8 Declaration of Decontamination................................... 19
3 Setup and Function.............................................................. 20
3.1 Basic Unit ..................................................................... 20
3.1.1 Main Switch .................................................. 21
3.1.2 LED Strip ...................................................... 21
3.1.3 Pumps .......................................................... 21
3.1.4 Identification Plate ........................................ 22
3.1.5 Power Connection ........................................ 23
3.1.6 Water Connections ....................................... 23
3.1.7 Gas Connections .......................................... 23
3.1.8 Signal Connections ...................................... 24
3.1.9 Motor Cable Connection ............................... 24
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3.1.10 Connections for Sensors (Sensor Cables) ...25
3.1.11 Sparger Connection (Gassing) .....................25
3.1.12 Connections and Water Flow Control Valve
for the Exit Gas Cooler .................................26
3.2 Operating Panel ...........................................................27
3.3 Culture Vessel ..............................................................27
3.3.1 Top Plate ......................................................29
3.3.2 Ports in the Vessel Top Plate and their
Configuration ................................................30
3.3.3 Vessel Top Plate, Nominal Width 90 ............30
3.3.4 Vessel Top Plate, Nominal Width 115 ..........31
3.3.5 Vessel Top Plate, Nominal Width 145 ..........32
3.4 Temperature Control System .......................................33
3.5 Stirrer ...........................................................................34
3.6 Gassing System ...........................................................35
3.6.1 Gas Entry ......................................................36
3.6.2 Exit Gas ........................................................36
3.7 pH Control ....................................................................36
3.7.1 Measurement System ...................................36
3.8 pO2 Control...................................................................37
3.8.1 Measurement System ...................................38
3.9 Antifoam Control ..........................................................39
3.9.1 Antifoam Sensor ...........................................39
4 Options ..................................................................................40
4.1 Turbidity Measurement ................................................40
4.1.1 Sensors .........................................................41
4.1.2 Calibrating the Zero Point .............................41
4.1.3 Mounting the Sensor ....................................42
4.1.4 Cleaning and Storing the Sensor ..................42
4.1.5 Interferences Turbidity Measurement ...........43
4.1.6 Specifications ................................................43
4.2 Exit Gas Analysis .........................................................44
4.2.1 Gas Sensors .................................................44
4.2.2 Connecting the Gas Sensor .........................44
4.2.3 Calibrating the Gas Sensor ..........................45
5 Accessories ..........................................................................46
5.1 Cone Plug for Drive Hub ..............................................46
5.2 Sparger ........................................................................47
5.3 Baffles ..........................................................................47
5.4 Blanking Plugs .............................................................48
5.5 Clamping Adapters and Fastening Screws ..................48
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5.6 Electrode Holder .......................................................... 49
5.7 Addition Port Adapters and Feed Needles .................. 50
5.8 Septum Collar .............................................................. 51
5.9 Dip Tubes ..................................................................... 51
5.10 Pocket for Temperature Sensor (Pt100) ...................... 52
5.11 Exit Gas Cooler ............................................................ 53
5.12 Reagent Bottles ........................................................... 54
5.13 Sampling System Super Safe Sampler ....................... 54
5.14 Pump Heads ................................................................ 57
5.15 Vessel Holder with Built-in Holder for Reagent Bottles
and Pumps ................................................................... 58
5.16 Sterile Filters ................................................................ 58
5.17 Hoses and Accessories ............................................... 60
5.18 O-Rings and Gaskets .................................................. 61
5.19 Inoculation Accessories and Tools .............................. 61
5.20 Starter Kit ..................................................................... 62
5.21 Service Sets ................................................................. 62
5.22 Auxiliary Supplies ........................................................ 62
6 Transport and Storage ........................................................ 63
6.1 Transport ...................................................................... 63
6.2 Storage ........................................................................ 64
7 Installation and Initial Operation ........................................ 65
7.1 General Location Requirements for Installation .......... 65
7.2 Minimum Distances ..................................................... 65
7.3 Connecting the Equipment to On-Site Supply Lines ... 65
7.3.1 Power Supply ............................................... 66
7.3.2 Water Supply and Return ............................. 66
7.3.3 Gas Supply ................................................... 67
7.3.4 Exit Gas ........................................................ 68
7.4 Connecting the Motor Cable ........................................ 68
7.5 Test Run ...................................................................... 69
7.5.1 Preparation Test Run ................................... 69
7.5.2 Cooling System ............................................ 70
7.5.3 Stirring .......................................................... 71
7.5.4 Heating and Adjusting Temperature ............ 71
7.5.5 Gassing ........................................................ 72
7.5.6 End of Test ................................................... 72
8 Before Cultivation ................................................................ 74
8.1 Preparing and Autoclaving the Culture Vessel ............ 74
8.1.1 Checking Gaskets (O-Rings) ........................ 74
8.1.2 Mounting the Impellers ................................. 75
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8.1.3 Mounting Dip Tubes and Spargers ...............76
8.1.4 Inserting the Vessel into the Vessel Holder ..77
8.1.5 Inserting the Baffles ......................................78
8.1.6 Moistening/Filling the Culture Vessel ...........79
8.1.7 Fitting the Vessel Top Plate ..........................79
8.1.8 Mounting the Blanking Plugs ........................80
8.1.9 Mounting Addition Port Adapters ..................81
8.1.10 Mounting the Feed Needle(s) .......................82
8.1.11 Mounting the Pocket for Temperature Sensor
(Pt100) ..........................................................82
8.1.12 Equipping the Port with a Septum Collar and
Septum for Inoculation ..................................82
8.1.13 Preparing the Dip Tube/Addition Port Adapter
for Inoculation ...............................................83
8.1.14 Mounting the Exit Gas Cooler .......................84
8.1.15 Preparing the Sensors ..................................85
8.1.16 Calibrating the pH Sensor ............................86
8.1.17 Mounting a Sensor into a 12 mm Port ..........86
8.1.18 Mounting Sensors with Electrode Holder .....87
8.1.19 Mounting the Antifoam Sensor .....................89
8.1.20 Preparing the Super Safe Sampler...............91
8.1.21 Mounting the Sparger Hose and the Inlet Air
Filter ..............................................................92
8.1.22 Preparing the Reagent Bottles, Pumps and
Hoses ............................................................93
8.1.23 Sterile Hose Connections .............................96
8.1.24 Setting the Pumps ........................................97
8.1.25 Removing the Pump Heads ..........................97
8.1.26 Fitting the Cone Plug for Drive Hub ..............98
8.1.27 Checklist Before Autoclaving ........................98
8.1.28 Autoclaving ...................................................99
8.2 Connecting the Culture Vessel and Preparing the
Cultivation ..................................................................101
8.2.1 Hang the Culture Vessel in Place and Fit the
Pump Heads ...............................................101
8.2.2 Filling the Reagent Hoses ..........................101
8.2.3 Connecting the Gassing .............................102
8.2.4 Connecting the Exit Gas Cooler .................102
8.2.5 Coupling the Motor .....................................103
8.2.6 Filling the Culture Vessel ............................104
8.2.7 Connecting the Temperature Sensor (Pt100)
....................................................................105
8.2.8 Connecting the Antifoam Sensor ................105
8.2.9 Connecting the pH Sensor .........................106
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8.2.10 Connecting the pO2 Sensor ........................ 106
8.2.11 Calibrating the pO2 Sensor ......................... 107
8.2.12 Checking the Hoses and Hose Connections
.................................................................... 107
9 Cultivation ........................................................................... 108
9.1 Preparing the Medium ............................................... 108
9.2 Sampling .................................................................... 109
9.3 Inoculation .................................................................. 112
9.3.1 Inoculation with a Syringe .......................... 113
9.3.2 Inoculation Using Dip Tube / Addition Port
Adapter ....................................................... 113
9.4 Harvest ....................................................................... 114
9.5 Emptying the Culture Vessel ..................................... 115
9.6 Emptying the Reagent Hoses .................................... 115
9.7 Switching off the Equipment ...................................... 115
9.8 Autoclaving the Culture Vessel After Cultivation ....... 116
10 Operation ............................................................................ 118
10.1 Screen Areas, Menu Navigation and Control Elements
................................................................................... 118
10.1.1 Main Screen ............................................... 120
10.1.2 EDIT VIEW ................................................. 121
10.1.3 START BATCH / INOCULATE / STOP
BATCH ....................................................... 122
10.1.4 SAMPLE NOW ........................................... 123
10.2 Menus for System Settings ........................................ 124
10.2.1 VESSEL TYPE – Selecting a Culture Vessel
.................................................................... 125
10.2.2 APPEARANCE – Display Settings ............. 126
10.2.3 NETWORK SETTINGS .............................. 128
10.2.4 eve COMMUNICATION – Communication
Settings ....................................................... 130
10.2.5 USB Data Export and Import from a USB
Stick ............................................................ 131
10.2.6 SYSTEM INFO – System Information ........ 134
10.3 Parameter - Parameter Groups ................................. 134
10.3.1 Parameters – Displays and Functions ....... 135
10.3.2 SETPOINT - Setting the Setpoint ............... 137
10.3.3 Parameter Alarms ...................................... 138
10.3.4 Cascades.................................................... 140
10.4 MAIN Parameter Group ............................................. 141
10.4.1 Temperature ............................................... 141
10.4.2 Stirrer .......................................................... 141
10.4.3 pH ............................................................... 141
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10.4.4 pO2 ..............................................................143
10.4.5 TotalFlow ....................................................146
10.4.6 GasMix ........................................................146
10.4.7 Foam ...........................................................147
10.5 EXTENDED Parameter Group ...................................149
10.5.1 Turbidity (Optional) .....................................149
10.5.2 Balance (Optional) ......................................149
10.5.3 AirFlow ........................................................149
10.5.4 Gas2Flow ....................................................150
10.6 EXITGAS Parameter Group .......................................151
10.6.1 Exit Gas O2 .................................................151
10.6.2 Exit Gas CO2 ..............................................152
10.7 PUMPS Parameter Group - General Information ......152
10.7.1 Configuring the Pumps ...............................154
10.7.2 Pump1 - Acid or Additional Feed Solution ..155
10.7.3 Pump2 - Base or Additional Feed Solution 156
10.7.4 Pump3 - Antifoam, Level or Additional Feed
Solution .......................................................156
10.7.5 Pump4 - Feed Solution ...............................157
10.7.6 AUTO FILL/EMPTY – Automatically
Filling/Emptying Pump Hoses .....................158
10.8 Calibration ..................................................................159
10.8.1 Calibrating the pH Sensor - General
Information ..................................................159
10.8.2 Calibrating the pH Sensor - Procedure ......160
10.8.3 pH Sensor Product Calibration ...................165
10.8.4 Calibrating the pO2 Sensor - General
Information ..................................................168
10.8.5 Calibrating the pO2 sensor - Procedure ......169
10.8.6 Calibrating the Turbidity Sensor - General
Information ..................................................176
10.8.7 Calibrating the Turbidity Sensor - Procedure
....................................................................176
10.9 PID Controller – Basic Principle .................................178
10.9.1 Table with Setting Values for PID Controller
....................................................................180
10.9.2 Useful Information for Changing PID
Controller Settings ......................................180
10.9.3 Adjusting PID Settings ................................181
10.10 Alarms – Equipment Alarm Menu ..............................181
11 Cleaning and Maintenance ................................................183
11.1 Cleaning Agent and Disinfectant ................................183
11.2 Cleaning the Culture Vessel - Routine Cleaning .......183
11.3 Removing the Vessel Top Plate and Accessories .....185
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11.3.1 Removing the Exit Gas Cooler ................... 185
11.3.2 Removing the Sensors ............................... 185
11.3.3 Removing Hoses, Filters and Pump Heads
.................................................................... 186
11.3.4 Removing Blanking Plugs .......................... 187
11.3.5 Removing the Septum Collar and Septum . 187
11.3.6 Removing Addition Port Adapters, Feed
Needle and Temperature Sensor Pocket ... 187
11.3.7 Removing the Vessel Top Plate ................. 187
11.3.8 Removing the Sparger and the Dip Tube(s)
.................................................................... 188
11.3.9 Removing the Impellers .............................. 189
11.4 Cleaning and Storing Individual Parts ....................... 189
11.5 Cleaning the Sensors ................................................ 190
11.6 Cleaning the Hoses and Pump Heads ...................... 190
11.7 Cleaning the Super Safe Sampler ............................. 191
11.8 Cleaning the Exit Gas Cooler .................................... 191
11.9 Cleaning the Basic Unit and Operating Panel ........... 192
11.10 Maintenance Plan ...................................................... 192
11.11 Decalcifying the Equipment ....................................... 193
12 Interferences ....................................................................... 195
12.1 Interferences Basic Unit and Operating Panel .......... 195
12.2 Interferences Drive System ....................................... 196
12.3 Interferences Temperature Control System .............. 197
12.4 Interferences Gassing System................................... 197
12.5 Interferences pH-System ........................................... 198
12.6 Interferences pO2 System .......................................... 200
12.7 Interferences Antifoam/Level Sensor and Antifoam
Pumps ........................................................................ 201
12.8 Interferences Addition of Feed Solution (Feed Pump)
................................................................................... 202
12.9 Returning for Repair .................................................. 202
13 Disassembly and Disposal ................................................ 203
13.1 Disassembly ............................................................... 203
13.2 Disposal ..................................................................... 203
14 Technical Data .................................................................... 205
14.1 Equipment Dimensions .............................................. 205
14.2 Dimensions of Culture Vessel.................................... 206
14.3 Weights (netto)........................................................... 207
14.4 Connection Requirements ......................................... 207
14.4.1 Electrical ..................................................... 207
14.4.2 Water IN, Basic Unit ................................... 207
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14.4.3 Water OUT, Basic Unit ...............................207
14.4.4 Gas (Air, O2 or N2) ......................................208
14.5 Specifications .............................................................208
14.5.1 Operating Panel ..........................................208
14.5.2 Culture Vessels...........................................208
14.5.3 Stirrer ..........................................................209
14.5.4 Temperature ...............................................210
14.5.5 Gassing .......................................................211
14.5.6 pH ...............................................................211
14.5.7 pO2 ..............................................................212
14.5.8 Antifoam ......................................................212
14.5.9 Pumps .........................................................213
14.6 Operating Conditions .................................................213
14.7 Emissions ...................................................................213
14.8 Auxiliary Supplies .......................................................214
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This operating manual is a component part of the equipment.
DANGER
will lead to severe or even fatal injuries if not avoided.
1 General Information
1.1 About this Manual
Minifors 2 - Benchtop Bioreactor
General Information
This manual enables the safe and efficient handling of the equipment.
All the information and instructions in this operating manual comply
with the current standards, legal regulations, the latest technological and scientific developments and the knowledge gained from the
manufacturer’s many years of experience in this field.
It must be kept near to the equipment and must be accessible
to the operators at all times.
The users must read the operating manual thoroughly and fully understand its contents before beginning any work.
Adhering to all the safety and operating instructions in this manual
is essential to ensure that work is carried out safely.
The scope of delivery may differ from the explanations, descriptions and figures in this operating manual due to special designs,
additional options specified on ordering and the latest technical/mechanical modifications.
This manual contains illustrations to aid general understanding.
These may differ from the actual equipment as supplied.
1.2 Explanation of Special Notices
1.2.1 Warning Notices
Warning notices in this manual are indicated by a coloured bar and
begin with a signal word that signifies the degree of the hazard.
The signal word “DANGER” indicates a dangerous situation that
Page 10 of 214 07 February 2019
WARNING
avoided.
CAUTION
situation that may result in minor injuries if not avoided.
ATTENTION
may result in significant damage to property if not avoided.
INFORMATION
efficient, fault-free operation of the equipment.
Minifors 2 - Benchtop Bioreactor
General Information
1.2.2 Other Notices
The signal word “WARNING” indicates a potentially dangerous
situation that may result in severe or even fatal injuries if not
The signal word “CAUTION” indicates a potentially dangerous
The word “ATTENTION” on a blue bar indicates a situation that
Texts located below a grey bar bearing the notice “INFORMATION” provide useful tips and recommendations for ensuring
1.3 Equipment Identification (Standard Identification Plate)
The identification plate is designed to allow clear identification of
the equipment. It contains the following information:
Manufacturer name
Designation = Category of equipment
Type = Equipment type (name)
S/N = Serial number
Year = Year of manufacture
Mains = Nominal voltage and frequency
Current = Current consumption
Manufacturer address
CE marking
07 February 2019 Page 11 of 214
1.4 Declaration of Conformity
The equipment is in compliance with the essential requirements of
the following Directives:
Machinery Directive 2006/42/EC
EMC Directive 2014/30/EU
The Declaration of Conformity according to EC Machinery Directive
2006/42/EC, annex II 1 A is included in the general documentation
supplied with the equipment.
1.5 Customer Service and Services
Minifors 2 - Benchtop Bioreactor
General Information
Our Customer Service is at your disposal for technical advice and
specialist enquiries. For contact information, see page 2.
Due to their familiarity with the potential applications of the equipment, the Customer Service team is able to provide information on
whether the equipment can be used for a specific application or
modified to handle the planned process.
Experience of working with the equipment will be published semiregularly on the manufacturer’s website in the form of “application
notes”.
Furthermore, our colleagues are always interested in new information and experiences resulting from user’s applications for the
equipment that may be valuable for the continued development of
our products.
Page 12 of 214 07 February 2019
It is essential to read the operating manual carefully – espe-
The bench-top bioreactor Minifors 2 from INFORS HT is de-
Minifors 2 - Benchtop Bioreactor
Safety and Responsibility
2 Safety and Responsibility
This section describes general considerations relating to user
safety that must be taken into account when working with the
equipment.
In the remaining sections, warning notices are used only to highlight particular hazards directly arising from the actions being described in the section in question.
cially this section and the warning notices in the text – and to
follow the instructions therein.
This section also refers to areas that are the responsibility of the
provider due to certain risks arising from particular applications for
which the equipment is used deliberately and with full awareness
of the associated risks.
2.1 Intended Use, Incorrect Use and Misuse
signed especially for research and development and for the
cultivation of microorganisms in a biotechnology laboratory.
The equipment is designed and constructed exclusively for the intended use described above.
Intended use also includes following all the instructions in this operating manual, especially those relating to:
The installation site
User qualifications
Correct operation and maintenance
The use of undamaged tubing and glass vessels
Any failure to observe the requirements specified in this manual
shall be deemed incorrect use.
Any use of the equipment outside the scope of the intended use as
described above shall be deemed misuse.
This also applies to applications for which the equipment is not designed, such as the use or production of explosive gases, which is
not permitted because the equipment is not explosion-proof.
07 February 2019 Page 13 of 214
2.2 Qualified Personnel
Minifors 2 - Benchtop Bioreactor
Safety and Responsibility
For use for special applications not covered by conventional, intended use, the equipment must be modified and certified accordingly by the manufacturer.
Any use of the equipment outside of a biotechnology laboratory,
i.e. in any environment in which the conditions required for the
safety of the users cannot be fulfilled or cannot be fulfilled to their
full extent, shall also be deemed misuse.
Due to the complexity of the equipment and the potential risks arising from its operation, the equipment may only be used by qualified, specialist personnel.
2.2.1 Provider
2.2.2 User
The term “provider” applies to all persons who are responsible for
making the equipment and the necessary infrastructure available.
These persons may also be included in the group of people known
as “users”, though this is not always the case.
Irrespective of whether a provider is a member of the company’s
board of management or a supervisor, they bear a special level of
responsibility with regard to the processes and the qualification and
safety of the users.
General
The term “user” applies to all persons who come into contact with
the equipment in any way and perform work on or with it. This primarily applies to the following activities, which can be performed by
the manufacturer’s own specialists or a variety of other persons (it
is not always possible to distinguish clearly between the different
types of person):
Assembly, installation and commissioning
Definition and preparation of the process
Operation
Troubleshooting and remedying of faults
Maintenance and cleaning (autoclaving, if necessary)
Service work and repairs
Disassembly, disposal and recycling
Page 14 of 214 07 February 2019
Minifors 2 - Benchtop Bioreactor
Safety and Responsibility
Qualified personnel
On account of their specific education, training and – in many
cases – experience, the qualified personnel required for this work
are able to recognise risks and respond accordingly to potential
hazards.
The qualified personnel (either internal or external) who cannot be
categorised under the separate “operators” group are made up of
the following groups of persons:
Electricians (electrical engineers)
Decontamination specialists
Repair specialists
Specialists in disassembly and (environmentally friendly) dis-
posal
Recycling specialists
2.2.3 Operator
The “operators” are a specific sub-group of users distinguished by
the fact that they work with the equipment. They are the true target
audience for this operating manual.
Qualified technicians
Only technicians who have been trained for working in a biotechnology laboratory can be considered for the role of operator. These
include:
Process technicians in the fields of biotechnology and chemis-
try
Biotechnologists (biotechnicians)
Chemists with a specialisation in biochemistry; chemists in the
field of organic chemistry or biochemistry
Life scientists (biologists) with special education in cytology,
bacteriology, molecular biology, genetics, etc.
Lab assistants (lab technicians) from various fields
In order to be classed as a “sufficiently qualified technician” for the
operation of the equipment, the persons in question must have received thorough training and have read and understood the operating manual.
The operator must be informed in a training session provided by
the provider of the tasks delegated to the operator and the potential risks of improper conduct. Tasks that go beyond the scope of
operation under normal conditions may only be performed by the
operator if this is specified in the manual and the provider has explicitly entrusted said tasks to the operator.
07 February 2019 Page 15 of 214
2.3 Unauthorised Persons
Minifors 2 - Benchtop Bioreactor
Safety and Responsibility
Technicians in training
Persons in this group who are undergoing training or apprenticeships are only permitted to use the equipment under supervision
and in accordance with the instructions of a trained and qualified
technician.
The term “unauthorised persons” applies to all persons who can
access the work area but are not qualified to use the equipment in
accordance with the aforementioned requirements.
Unauthorised persons are not permitted to operate the equipment
or use it in any other way.
2.4 Responsibility of the Provider
The equipment is used for industrial and scientific purposes. As
such, the provider of the equipment is individually liable with regard
to the legal requirements relating to occupational health and safety
in a biotechnology laboratory. In particular:
The provider is responsible for ensuring that the work and en-
vironmental regulations applicable in a biotechnology laboratory are observed.
The provider must ensure that the equipment remains in safe
and proper working condition throughout its entire term of use.
The provider must ensure that all safety equipment is fully
functional and is not disabled.
The provider must ensure that the equipment is only worked
on by qualified users, and that said users receive sufficient
training.
The provider must ensure that the protective equipment re-
quired for working with the equipment is provided and worn.
The provider must ensure that this operating manual remains
in the immediate vicinity of the equipment throughout its entire
term of use.
2.5 General Hazards
This section covers general hazards and residual risks that are always present when using the equipment in accordance with normal, intended use.
Page 16 of 214 07 February 2019
The equipment runs on electrical power. There is an immediate
facturer represent a significant safety risk. As such, we recommend
Minifors 2 - Benchtop Bioreactor
Safety and Responsibility
2.5.1 Electrical Current
The following notices are general in nature. As such, with a few
exceptions they are not repeated in the remaining sections.
risk of fatal injury if contact is made with live parts.
The following points must be observed in order to avoid the risk of
fatal injury:
In case of damage to insulation, disconnect the equipment
from the mains immediately and arrange for it to be repaired.
Disconnect the equipment from the mains before commencing
any work on the electrical system.
Always use qualified electricians for any work on the electrical
system.
Keep moisture away from live parts. It may lead to a short cir-
cuit.
2.5.2 Unauthorised Spare Parts and Accessories
Incorrect or imitated spare parts and accessories as well as spare
parts or accessories that have not been authorised by the manu-
procuring all spare parts and accessories from an authorised
dealer or directly from the manufacturer. For the contact details of
the manufacturer’s representatives, see page 2.
2.6 Particular Hazards
This section covers particular hazards and residual risks that may
arise when using the equipment for special applications in accordance with normal, intended use.
Since the use of the equipment for such applications is deliberate,
it is the responsibility of the operators and the provider to ensure
that all personnel are protected from potential damage to health.
The provider is responsible for ensuring that the appropriate protective equipment for such applications is provided, and that the
necessary infrastructure is in place.
07 February 2019 Page 17 of 214
For processes that are carried out with temperatures over 55 °C,
The use or production of dangerous gases i.e. toxic or asphyxiant
The use or production of flammable or explosive substances is not
The use or production of corrosive or toxic substances entails a
2.6.1 Hot Surfaces
2.6.2 Dangerous Gases
Minifors 2 - Benchtop Bioreactor
Safety and Responsibility
there is a danger of burns on hot surfaces.
Since the equipment is intended for applications at high temperatures, it is the responsibility of the users to ensure that they have
sufficient protection.
The motor gets hot during operation. There is a risk of burns if it is
touched.
The thermal block and its adapter get hot during operation. There
is a risk of burns, if touched.
gases entails a significant health risk, especially in enclosed
spaces.
In order to prevent high emissions of dangerous gases, the following measures must be taken:
The gas connections on the equipment must be checked be-
fore any cultivation processes using dangerous gases are initiated.
The gaskets on the equipment must be checked at regular
intervals and replaced if necessary.
Siphon off exit gas safely.
2.6.3 Flammable or Explosive Substances
covered under “intended use” of the equipment, as the equipment
is not explosion-proof.
If the provider intends to use the equipment for such purposes, he
must check its suitability for the planned application with the responsible local authorities.
2.6.4 Corrosive or Toxic Substances
significant health risk. As such, special measures must be taken to
protect the users for such applications.
Page 18 of 214 07 February 2019
The use or production of bioactive substances, pathogenic organGlass vessels may break or shatter when subjected to overpres-
WARNING
with warning symbols on the equipment are always intact.
Minifors 2 - Benchtop Bioreactor
Safety and Responsibility
Since the equipment is used deliberately for such applications, it is
the responsibility of the users to ensure that they have sufficient
protection.
2.6.5 Bioactive Substances or Pathogenic Organisms
isms or genetically modified cultures entails a significant health
2.6.6 Overpressure or Vacuum
risk. As such, special measures must be taken to protect the users
for such applications.
Since the equipment is used deliberately for such applications, it is
the responsibility of the users to ensure that they have sufficient
protection.
sure or vacuums.
2.7 Warning Symbols on the Equipment
The following warning symbols (stickers) are attached to the equipment:
Position
Thermal block adapter
Motor
Illegible or missing warning symbols on the equipment will lead
to the user being exposed to risks that the warning symbols in
question were designed to make him or her aware of.
It is the provider’s responsibility to ensure that all the stickers
07 February 2019 Page 19 of 214
2.8 Declaration of Decontamination
When returning the equipment for repair, disassembly or disposal,
it is required for the safety of all parties involved and because of
legal provisions that a lawful declaration of decontamination is present.
The following must be observed if this is the case:
The equipment, the component part or accessory must be en-
tirely decontaminated before sending to the manufacturer
The provider is therefore required to completely and truthfully
fill out a declaration of decontamination, and have it signed by
the person responsible.
The declaration of decontamination must be affixed on
the outer packaging in which the equipment is sent back.
These forms can be obtained from the licensed dealer or the
manufacturer. See address on page 2.
Important notice
If the return shipment is not accompanied by a signed and complete declaration of decontamination or it is not affixed to the outer
packaging, the shipment will be returned unopened to the sender
at their expense (see also T&C).
Minifors 2 - Benchtop Bioreactor
Safety and Responsibility
Page 20 of 214 07 February 2019
Minifors 2 - Benchtop Bioreactor
Setup and Function
3 Setup and Function
3.1 Basic Unit
4
5
2
3
6
1
7
8
1 Pumps
2 Thermal block and adapter
3 Connections for sparger and exit gas cooler
4 Operating panel
5 LED signal strip
6 Main switch
7 Connections for sensors
8 Hooks for vessel holder
All of the measurement and control technology is built into the
basic unit. The basic unit is equipped as standard with a thermal
07 February 2019 Page 21 of 214
The main switch is located on the right-hand side on the basic unit.
3.1.1 Main Switch
Minifors 2 - Benchtop Bioreactor
Setup and Function
block plus adapter for regulating the temperature of the culture
vessel, four pumps for adding reagents and feed solution, and the
operating panel.
It lights up green as soon as it is switched on.
3.1.2 LED Strip
The LED strip is located on the front of the basic unit.
When the LED strip:
Lights up green as soon as the equipment is switched on.
The equipment is functioning as normal.
Flashes green. This indicates a parameter alarm (for more
details, see the “Operation” section).
Flashes red. This indicates an Equipment error (for more de-
tails, see the “Interferences” section).
3.1.3 Pumps
Reagents and feed solution are supplied via four peristaltic pumps.
The pumps are driven by stepper motors.
Page 22 of 214 07 February 2019
The pump drive shafts are located on the left-hand side of the
The autoclavable pump heads are plugged into a mounting plate.
Minifors 2 - Benchtop Bioreactor
Setup and Function
basic unit. The drive shafts’ direction of rotation is set as standard
to anti-clockwise for “filling”; see marking on mounting plate. The
pumps can be configured individually using the operating panel,
and thus each set to digital or analogue operation as required.
A hinged Plexiglas cover acts as a guard during operation.
(Depicted separately here in order to show the marking under the
pumps.) This is numbered 1 to 4 from bottom to top, and labelled
to indicate the standard factory settings:
Pump 1: Acid (digital)
Alternative setting: FEED (analogue)
Pump 2: Base (digital)
Alternative setting: FEED (analogue)
Pump 3: AF (antifoam, digital)
Alternative setting: LEVEL (digital) or FEED (analogue)
Pump 4: Feed (analogue)
Alternative setting: BALANCED (analogue)
For more information on possible pump settings, see the “Operation” section, sub-section “Parameter Group PUMPS”.
The pump heads and the mounting plate can be simply pushed
onto or pulled off the drive shafts.
3.1.4 Identification Plate
The identification plate is located at the side of the basic equipment.
The data provided on the identification plate is specified in the
main chapter “General Information”, chapter “Equipment Identification”.
07 February 2019 Page 23 of 214
The connection socket for the power cable is located on the rear of
3.1.5 Power Connection
3.1.6 Water Connections
Minifors 2 - Benchtop Bioreactor
Setup and Function
the basic unit, at the bottom left.
3.1.7 Gas Connections
The water connections are located on the rear of the basic unit, at
the bottom right. They are marked with the following symbols:
H
O IN: water inlet
2
H
O OUT: water outlet
2
The connections for the gas supply are located on the rear of the
basic unit, at the bottom right, above the water connections.They
are marked with the following symbols:
Inlet oxygen / nitrogen
Inlet air
Page 24 of 214 07 February 2019
The connection for the motor cable is located on the rear of the
Minifors 2 - Benchtop Bioreactor
Setup and Function
3.1.8 Signal Connections
The following signal connections with the corresponding symbols
are located on the rear of the basic unit, at the bottom left:
SERVICE: 9-pin RS232, for connecting a diagnostic com-
puter for maintenance.
BALANCE: 9-pin RS232, for connecting a balance
LAN port: for connecting a network cable
3.1.9 Motor Cable Connection
basic unit at the top left, and marked with a corresponding symbol.
07 February 2019 Page 25 of 214
The basic unit is equipped as standard for measuring temperature,
The connection for the gassing (sparger) is located on the front of
1
2
3
4
5
3.1.10 Connections for Sensors (Sensor Cables)
pH, pO2 and antifoam. This means that the temperature (Pt100)
and the connecting cables for the pH, pO
are always present. The matching sensors for pH, pO
foam are included in the standard package.
1 pO
2 Temperature (Pt100)
3 Antifoam
4 pH
5 Spare connection for turbidity measurement sensor (option)
2
Minifors 2 - Benchtop Bioreactor
Setup and Function
and antifoam sensors
2
and anti-
2
3.1.11 Sparger Connection (Gassing)
the basic unit, at the bottom left. It is marked with a corresponding
symbol.
Page 26 of 214 07 February 2019
The hose used to connect the sparger is connected to the basic
The water connections for the exit gas cooler are located on the
The water supply and return hoses for the exit gas cooler are con-
INFORMATION
them to the exit gas cooler incorrectly.
Minifors 2 - Benchtop Bioreactor
Setup and Function
unit’s gassing connection at the factory.
3.1.12 Connections and Water Flow Control Valve for the Exit Gas Cooler
front of the basic unit, at the bottom left. They are marked with the
corresponding symbols:
1 Exit gas cooler water outlet
2 Exit gas cooler water inlet
1
2
nected to the basic unit in the factory.The rapid couplings at both
ends of the hoses are used to connect them to the exit gas cooler.
Due to the different hose lengths, it is not possible to connect
07 February 2019 Page 27 of 214
The water flow control valve is located on the rear of the basic unit
The operating panel on the top right of the basic unit has a 7” TFT
3.2 Operating Panel
Minifors 2 - Benchtop Bioreactor
Setup and Function
and marked with a corresponding symbol:
The control valve is set at the factory. If necessary, the water flow
rate can be set manually using the control valve.
Turn anti-clockwise to increase the water flow rate
Turn clockwise to reduce the flow rate
The valve can be fixed in the required position using a lock nut.
colour touch screen.
On the right-hand side of the panel is a USB port.
On the left-hand side is a slot for an SD card
The operating panel is switched on using the main switch. For a
detailed description of how to operate it, see the “Operation” section.
3.3 Culture Vessel
The culture vessel comprises a glass vessel, the top plate with the
standard fittings (which vary based on vessel size) and the vessel
holder with handles. The vessel is made of borosilicate glass.
Page 28 of 214 07 February 2019
Minifors 2 - Benchtop Bioreactor
Setup and Function
8
1
6
5
7
2
3
4
1 Motor coupling
2 Vessel holder handle
3 Glass vessel
4 Vessel holder stand
5 Pump holder
6 Reagent bottle holder
7 Vessel holder
8 Top plate
The illustration shows a culture vessel with a total volume of 1.5 L
and a nominal width of 90 mm. There are three vessel sizes available, each with a matching top plate.
The vessel holder has two handles on the side, which are used
when emptying and cleaning the vessel or transporting it to the autoclave.
07 February 2019 Page 29 of 214
3.3.1 Top Plate
Minifors 2 - Benchtop Bioreactor
Setup and Function
1
2
3
1 Knurled screw (x4)
2 Top plate
3 Vessel
4
5
6
4 O-ring
5 Damping ring (spacer ring)
6 Flange
The top plate is attached to the vessel using four knurled screws
and a flange. The knurled screws also hold the vessel in place in
the vessel holder. An O-ring is used to seal the top plate. A spacer
ring is used to prevent the top plate from exerting pressure on the
rim of the vessel.
Page 30 of 214 07 February 2019
Minifors 2 - Benchtop Bioreactor
Setup and Function
3.3.2 Ports in the Vessel Top Plate and their Configuration
The vessel top plate has different ports of different sizes to mount
the different components such as sparger, blanking plugs, sensors
etc. The number of ports in the top plate and its configuration depends on the diameter nominal (= inner diameter) of the culture
vessel.
3.3.3 Vessel Top Plate, Nominal Width 90
1
2
6
7
8
3
9
4
10
5
1 Ø 12 mm Pg13.5: pH sensor
2 Ø 12 mm Pg13.5: Exit gas cooler
3 Ø 7.5 mm: Addition port adaptors (4x)
4 Ø 10 mm: Sparger
5 Ø 10 mm: Dip tube for sampling
6 Ø 12 mm Pg13.5: pO2 sensor
7 Ø 10 mm: Temperature sensor (Pt100)
8 Antifoam sensor earth connection
9 Ø 10 mm: Antifoam sensor
10 Ø 12 mm Pg13: Inoculation
07 February 2019 Page 31 of 214
2 3 4
5
1
6
7
8
9
10
11
12
3.3.4 Vessel Top Plate, Nominal Width 115
Minifors 2 - Benchtop Bioreactor
Setup and Function
1 Ø 12 mm Pg13.5: pH sensor
2 Ø 12 mm Pg13.5: Exit gas cooler
3 Ø 12 mm Pg13.5: Additional sensor
4 Ø 7.5 mm: Addition port adaptors (4x)
5 Ø 10 mm: Sparger
6 Ø 12 mm Pg13.5: pO
sensor
2
7 Ø 12 mm Pg13.5: Additional sensor
8 Ø 10 mm: Temperature sensor (Pt100)
9 Antifoam sensor earth connection
10 Ø 10 mm: Antifoam sensor
11 Ø 12 mm Pg13.5: Inoculation
12 Ø 10 mm: Dip tube for sampling
Page 32 of 214 07 February 2019
Minifors 2 - Benchtop Bioreactor
Setup and Function
3.3.5 Vessel Top Plate, Nominal Width 145
1
2
3
6
7
8
9
10
4
11
12
5
13
1 Ø 12 mm Pg13.5: Exit gas cooler
2 Ø 12 mm Pg13.5: Additional sensor
3 Ø 7.5 mm: Addition port adaptors (4x)
4 Ø 10 mm: Sparger
5 Ø 10 mm: Dip tube for sampling
6 Ø 12 mm Pg13.5: pH sensor
7 Ø 12 mm Pg13.5: pO
sensor
2
8 Ø 10 mm: Temperature sensor (Pt100)
9 Antifoam sensor earth connection
10 Ø 10 mm: Antifoam sensor
11 Ø 12 mm Pg13.5: Additional sensor
12 Ø 12 mm Pg13.5: Additional sensor
13 Ø 12 mm Pg13.5: Inoculation
07 February 2019 Page 33 of 214
The two hooks on the thermal block hold the culture vessel in place
3.4 Temperature Control System
The temperature (heating and cooling) is controlled using a thermal
block and its adapter.
1 Thermal block adapter
1
2
2 Thermal block
3 Fastening screw (Allen screw, 4 pieces)
4 Hook, 2 pieces
There is a thermal block adapter for each vessel size. The thermal
block adapters are screwed onto the thermal block.
Minifors 2 - Benchtop Bioreactor
Setup and Function
3
4
The temperature in the culture vessel is measured using a platinum resistor temperature sensor (Pt100). The temperature is
transmitted from the thermal block to the adapter and from the
adapter to the culture vessel by means of heat exchange.
The thermal block is heated electrically using heating cartridges. It
is cooled by allowing cooling liquid to flow through it.
on the basic unit. In order to ensure optimum heat transmission,
the two hooks also pull the culture vessel right up against the thermal block.
Page 34 of 214 07 February 2019
The stirrer shaft is driven from above and turns anti-clockwise (left
Minifors 2 - Benchtop Bioreactor
Setup and Function
3.5 Stirrer
1
when viewing vessel from above).
1 Drive hub
2 Stirrer shaft
3 Mechanical seal
The stirrer shaft is sealed using a mechanical seal.
2
3
Two Rushton impellers are attached to the stirrer shaft by means
of grub screws.
07 February 2019 Page 35 of 214
A brushless gear motor with a mechanical coupling is used as
The motor is connected by pushing it onto the drive hub on the top
Minifors 2 - Benchtop Bioreactor
Setup and Function
standard. Depending on the size of the vessel, two motors with different power levels are used; see the “Technical data” section,
sub-section “Stirrer specifications”.
Left: Small motor for culture vessels with nominal width 90
Right: Large motor for culture vessels with nominal widths
115 and 145
plate.
3.6 Gassing System
The following gases can be used:
Air
Oxygen (O
The basic unit is equipped and configured with two mass flow controllers for controlling the gas flow and, if necessary, the gas mixture. If oxygen or nitrogen are used in addition to air, the gases are
mixed before being fed into the culture vessel. Both the gas flow
rate(s) and the composition of the gas mixture (where applicable)
are set using the operating panel. For more details, see the “Operation” section.
) or nitrogen (N2)
2
Page 36 of 214 07 February 2019
INFORMATION
installed upstream of the exit gas filter as a foam trap.
Minifors 2 - Benchtop Bioreactor
Setup and Function
3.6.1 Gas Entry
3.6.2 Exit Gas
A silicone hose leads the gas or gas mixture from the gassing connection on the basic unit to the culture vessel, via a sterile filter.
The gas is fed directly into the medium via the sparger (sparger
gassing). For more details on the sparger, see the “Accessories”
section, sub-section “Sparger”.
Even without active gassing, any cultivation can increase the pressure inside the vessel through heating or gas production. As such,
an exit gas line is essential for all cultivation processes.
Siphoning off exit gas via the exit gas cooler
The exit gas cooler dries the exit gas through condensation, thus
preventing the exit gas filter from becoming clogged with moisture.
At the same time, it also prevents liquid loss in the culture medium.
3.7 pH Control
3.7.1 Measurement System
If heavy foaming is expected, a bottle of antifoam agent can be
The exit gas cooler is included in the standard package; for more
details, see the “Accessories” section, sub-section “Exit Gas
Cooler”.
pH in the medium is measured by the pH sensor and regulated by
addition of reagents (acid, base). Addition of acid and base takes
place via the two peristaltic pumps Acid and Base .
Reagent bottles are filled with acid and base which are connected
to an addition port adapter in the vessel top plate and the two
pumps by silicone hoses.
The pH measurement system is designed for use with HAMILTON
digital sensors:
Conventional pH sensor (potential measurement against refer-
ence) with built-in electronics
07 February 2019 Page 37 of 214
INFORMATION
before use.
INFORMATION
the operating panel is no longer necessary.
Minifors 2 - Benchtop Bioreactor
Setup and Function
Type: Easyferm Plus ARC
Manufacturer: HAMILTON
The matching pH sensor is included in the standard package.
These pH sensors are pre-configured by the equipment manufacturer, INFORS HT. Replacement sensors must be configured
For details on the technical data, maintenance and storage requirements for the pH sensors, see the separate documentation provided by the sensor manufacturer. Read and follow the instructions.
Calibration of the pH sensor is always carried out BEFORE autoclaving. This is done on the operating panel. For more details on
this procedure refer to the main chapter “Operation”, chapter “Calibrating the pH Sensor”.
3.8 pO2 Control
If the pH sensor has already been calibrated before connection
to the system, the bioreactor will use this data and calibration on
For culture vessels with nominal widths of 90 and 145, pH sensors
can be mounted directly into 12 mm/Pg13.5 ports. For culture vessels with a nominal width of 115, an electrode holder is used.
For more details on the electrode holder, see the “Accessories”
section, sub-section “Electrode Holder”.
The oxygen saturation of the (culture) medium is measured by the
pO
sensor, and can be adjusted as follows:
2
Increasing the pO
The content of the oxygen dissolved in the medium (pO
2
) can be
2
increased using the following methods:
Increasing the stirrer speed
Increasing the gas volume flow rate (air and/or oxygen)
Increasing the oxygen content in the Gasmix.
Page 38 of 214 07 February 2019
INFORMATION
configured before use.
Minifors 2 - Benchtop Bioreactor
Setup and Function
3.8.1 Measurement System
These approaches can also be combined.
reduction
pO
2
In anaerobic processes, the vessel can be gassed using nitrogen.
This displaces the oxygen dissolved in the medium.
The pO2 measurement system is designed for use with HAMILTON
digital sensors:
pO
Type: Visiferm DO ARC
Manufacturer: HAMILTON
sensor with built-in opto-electronics
2
The HAMILTON pO2 sensors are pre-configured by the equipment manufacturer, INFORS HT. Replacement sensors must be
For details on the technical data, maintenance and storage requirements for the pO
sensors, see the separate documentation pro-
2
vided by the sensor manufacturer. Read and follow the instructions.
Generally speaking, the following applies: Unlike measurements
such as pH, which are calibrated to absolute measurement values,
the oxygen measurement is always calibrated to a relative reference point. For this purpose, the calibration is set to 100 % relative
oxygen saturation, usually with air at max. stirring speed and maximum gas flow rate. The actual concentration of dissolved oxygen
in mmol/L may therefore vary at 100 % saturation, depending on
the process.
Calibration is always carried out AFTER autoclaving. This is done
using the operating panel. Depending on the specifications defined
by the user, the pO
sensor will be calibrated either before the ves-
2
sel is filled with medium or afterwards, in the prepared medium.
For more details on this procedure refer to main chapter “Operation”, chapter “Calibrating the pO
Sensor”.
2
For culture vessels with nominal widths of 90 and 145, pO
sen-
2
sors can be mounted directly into 12 mm/Pg13.5 ports. For culture
vessels with a nominal width of 115, an electrode holder is used.
07 February 2019 Page 39 of 214
Inside diameter
2 mm
Hose connection outside diame-
4 mm
ring is used to mount the sensor
3.9 Antifoam Control
Minifors 2 - Benchtop Bioreactor
Setup and Function
For more details on the electrode holder, see the “Accessories”
section, sub-section “Electrode Holder”.
Foam hinders the exchange of gas between the medium and the
gas phase in the head space. The exit gas filter can become
clogged with foam, which causes a pressure build-up in the vessel.
This can be prevented by adding antifoam agent.
The antifoam agent is kept in a reagent bottle that is connected to
the antifoam sensor and the antifoam pump via a hose. The sensor
also acts as a dosing needle. When the sensor comes in contact
with foam, the antifoam pump is activated and antifoam agent is
fed into the vessel via the dosing needle.
3.9.1 Antifoam Sensor
A
B
1
ter
A clamping adapter with a fixed Oin the 10 mm port in the vessel top plate.
2
1 Sensor head with port for banana connector (A)
2 Clamping adapter with slotted-head screw (B)
3 Needle with transparent insulation
3
The antifoam sensor is equipped with two NON-autoclavable protective caps.
Page 40 of 214 07 February 2019
INFORMATION
eas.
Minifors 2 - Benchtop Bioreactor
Options
4 Options
4.1 Turbidity Measurement
The following options are available in addition to the equipment
included in the scope of supply for the basic unit.
The Optek ASD12-N measurement sensor can be used to determine the turbidity of the culture. Turbidity measurement can be
used to draw conclusions regarding the biomass concentration in
the culture.
The system comprises a sensor with a transmitter that is integrated
into the bioreactor basic unit:
Type ASD12-N with OPL05 (optical path length) for higher cell
densities.
Manufacturer: Optek
Measures the absorption in the 0 – 4 CU range
The ASD12-N sensors supply a non-linearised turbidity measurement for the culture. This can be linearised manually using the soft
®
sensor in eve
, for example, in order to determine correlation with
factors such as the biomass concentration or optical density.
The Optek ASD sensors are designed exclusively for use as turbidity sensors for liquids and gases in accordance with the technical data (“Specifications”).
The use of these sensors is forbidden in potentially explosive ar-
07 February 2019 Page 41 of 214
The sensors possess a measurement segment that is optimised in
he media
4.1.1 Sensors
Minifors 2 - Benchtop Bioreactor
Options
Optek ASD sensors are high-precision turbidity sensors. They can
be used to draw conclusions regarding the growth of microbial cultures or cell cultures as a function of NIR absorption.
1
2
1 Measurement gap (optical path length, OPL)
2 Shaft, Ø 12 mm
3 PG 13.5
terms of flow and sterility. The seal-free design of the sapphire window means that there are no gaps or joints. This guarantees complete sterility. All components that come into contact with t
are made of electrolytically polished steel.
1
2
3
4
5
Main components
1 LED light source
2 Sapphire window
3 Optical path length (OPL)
4 Daylight filter
5 Silicon photodiode detector
A defined LED light beam is shone through the process medium.
The weakening of the light intensity, which is caused by absorption
and/or diffusion by the dissolved and undissolved particles in the
carrier medium, is detected by a hermetically encapsulated silicon
photodiode.
The ASD-N sensors use the light in the near-infrared range (NIR)
of 840 nm to 910 nm.
3
4
4 Sensor head
5 Connection (push-pull plug connection)
5
4.1.2 Calibrating the Zero Point
Optek sensors are pre-calibrated in the factory. Inserts are available for reference measurement.
Due to the different light absorption of different media, zero point
calibration should be performed before each cultivation process.
Page 42 of 214 07 February 2019
Minifors 2 - Benchtop Bioreactor
Options
4.1.3 Mounting the Sensor
This can be done on the operating panel, either before or after
autoclaving, depending on the application in question. For more
details, see the main chapter “Operation” chapter “Calibrating the
Turbidity Sensor”.
For culture vessels with nominal widths of 90 and 145, Optek ASD
sensors can be mounted directly into 12 mm/Pg13.5 ports. For culture vessels with a nominal width of 115, an electrode holder is
used. For more details on the electrode holder, see the “Accessories” section, sub-section “Electrode Holder”.
Please note the following points when mounting the components:
Ensure that the sensor is fitted with an O-ring; fit an O-ring if
necessary.
Mount the sensor by hand – do not use any tools!
If the mounting depth of the sensor is adjustable (mounting
with electrode holder), make sure the mounting depth is set
correctly prior to autoclaving, as later adjustment represents a
contamination risk.
Mount the sensor in such a way that it cannot come in contact
with other components or the glass vessel.
Mount the sensor in such a way that it has good access to the
flow and there is no risk of bubbles collecting in the measurement gap.
4.1.4 Cleaning and Storing the Sensor
A build-up of deposits on the sapphire window will cause shifts in
the zero point and restrict the dynamic range of the sensor. As
such, the sapphire window on the sensor must be checked for deposits before each use and cleaned if necessary.
If necessary, store the Optek sensor in a clean, dry place at a temperature of between -20°C and 70°C.
07 February 2019 Page 43 of 214
Interference
Displayed measured value is not plausible / unusual
Possible Cause
Remedy
By
Sensor cable is twisted or kinked or
Check and ensure that the sensor cable is not
kinked or twisted. Connect the sensor cable properly
Operator
Sensor is not calibrated
Calbrate the zero point
Operator
Window fouling on the sapphire win-
Carefully clean the sensor
Operator
Qualified elecFaulty sensor
Replace the whole sensor
Operator
INFORMATION
measurement automatically continues.
Description
Value
Sensor type and
ASD12-N
Measuring principle
1-channel absorption of light
Measurement wave-
840 nm – 910 nm
Detector
Silicone photodiode (hermetically sealed)
4.1.5 Interferences Turbidity Measurement
not properly connected.
as necessary.
dows .
Faulty sensor cable Replace sensor cable
Minifors 2 - Benchtop Bioreactor
Options
trician
4.1.6 Specifications
The measuring system automatically switches off, if the temperature of the sensor in the medium exceeds 50 °C during operation (thermal shutdown). Once the medium has cooled down,
manufacturer
Path length OPL05: for higher cell densities
Material sensor 316L, electropolished and sapphire glass
length
Measuring range 0 – 4 CU (concentration units)
Protection IP 68
Light source Hybride LED
Temperature Shutdown at 50 °C1
1)
The measuring system automatically switches off, if the temperature
of the sensor in the medium exceeds 50 °C during operation (thermal
shutdown). Once the medium has cooled down, measurement automatically continues.
Page 44 of 214 07 February 2019
Vol. % O2
Vol. % CO2
1.0 – 50
0 – 10
or
0 – 25
INFORMATION
cesses!
Minifors 2 - Benchtop Bioreactor
Options
4.2 Exit Gas Analysis
4.2.1 Gas Sensors
In order to allow the user to draw conclusions regarding the status
of the culture while the bioprocess is still underway, the CO
O
measurements are often taken and analysed in the exit gas flow
2
and
2
of the bioreactor.
Combined CO2 and O2 gas sensors of the type BlueInOneFerm or
Blue Vary from the manufacturer BlueSens are available for exit
gas analysis.
3 m of pressure hose, D = 8 x 14.5 and a clamp are included for
connecting the gas sensor to the culture vessel (exit gas filter).
Measurement ranges gas sensors
Both sensor types are only suitable for use in aerobic bopro-
For details on the safety, technical data, usage and maintenance
requirements for the gas sensors, see the separate documentation
provided by the sensor manufacturer. Read this documentation
before using the gas sensor and follow the instructions contained
therein.
4.2.2 Connecting the Gas Sensor
In order to view measurements on the operating panel, you must
connect the gas sensor to the sensor cable and channel the exit
gas from the bioreactor through the gas sensor using a hose. The
cable is usually connected once during commissioning and can
remain untouched thereafter. The connection to the exit gas line
must be re-established before each cultivation process.
The ideal connection conditions are detailed in the separate documentation provided by the manufacturer.
07 February 2019 Page 45 of 214
INFORMATION
nect at this point, e.g. for autoclaving the culture vessel.
Minifors 2 - Benchtop Bioreactor
Options
Connecting the sensor cable
The fixed sensor cable is pre-installed in the factory (rear of equipment). The cable has an 8-pin round plug connector. In order to
connect the sensor, the plug connector is plugged into the socket
marked Port A on the gas sensor.
Due to the length of the sensor cable, the gas sensor can be positioned in a large number of possible locations.
Establishing the hose connection
The hose connection between the culture vessel (exit gas filter)
and the gas sensor must be designed in line with the direction in
which the gas flows through the gas sensor.
Proceed as follows:
Procedure 1. Cut as short a piece as possible off the supplied pressure
hose.
2. Push one end of the hose onto the hose nozzle (observe di-
rection of flow) on the gas sensor’s flow adapter and fasten in
place with the clamp.
3. Push the open end of the hose onto the exit gas filter on the
exit gas cooler.
Do NOT use a clamp here, as the hose must be easy to discon-
4.2.3 Calibrating the Gas Sensor
1-point calibration must be carried out once per month and during
initial commissioning in order to guarantee exact measurement results.
This is done directly on the gas sensor itself. The procedure is described in the separate documentation provided by BlueSens.
4.2.1 Replacing the BlueVary Gas Sensor Cartridge
The max. operating time of a BlueVary gas sensor cartridge
amounts to 9000 operating hours. Once this limit is reached, measurement is no longer possible. I.e. there is no measured value output anymore and the display turns red. The gas sensor cartridge
must be replaced by the sensor manufacturer.
For detailed information refer to the separate documentation from
the sensor manufacturer.
Page 46 of 214 07 February 2019
Accessories
1.5 L TV/NW 90
3.0 L TV/NW 115
6.0 L TV/NW 145
Impeller, Rushton
2 2 2
Dip tube, straight, Ø 6 mm for
1 1 1
Antifoam sensor for 10 mm port
1 1 1
Blanking plug for 12 mm/Pg13.5 port
4 6 7
Blanking plug for 10 mm port
2 2 2
Exit gas cooler for 12 mm/Pg13.5 port
1 1 1
Reagent bottle, 250 mL
4 4 4
Pump heads with hoses
1.1
4 4 4
pO2 sensor, Visiferm DO
1 1 1
pH sensor, Easyferm
1 1 1
Starter kit 1 1
1
Electrode holder for 12 mm/Pg13.5 port
2 2 2
The cone plug (EPDM) provided in the starter set protects the drive
hub from penetration of condensate water during sterilisation in the
Minifors 2 - Benchtop Bioreactor
Accessories
5 Accessories
The table below lists all the accessories included in the standard
package, divided according to vessel size (TV = total volume) and
nominal vessel width (inside diameter).
Baffles 1 1 1
Sparger, ring-shaped 1 1 1
Pocket for temperature sensor
in 10 mm port
12 mm/Pg13.5 port
1 1 1
Addition port adaptor, for 7.5 mm port 4 4 4
Clamping adapter for 10 mm port 3 3 3
Inside diameter: 1.0 mm/wall thickness:
mm
Cone plug for drive hub 1 1 1
5.1 Cone Plug for Drive Hub
autoclave.
07 February 2019 Page 47 of 214
It must be plugged into the opening of the drive hub for autoclaving
Inside diameter
4.0 mm
Hose connection outside diame-
6.0 mm
The baffles are used to mix the culture. They are simply inserted
5.2 Sparger
Minifors 2 - Benchtop Bioreactor
Accessories
of the culture vessel!
5.3 Baffles
The gas is fed directly into the medium via a ring sparger (Ø 6
mm). The sparger is mounted in a 10 mm port in the vessel top
plate using a clamping adaptor, and connected to the gassing system on the basic unit via a silicone hose with a sterile filter.
ter
into the glass vessel.
Page 48 of 214 07 February 2019
Fitted with fixed O-ring.
A fastening screw is used to fasten it in the 10 mm vessel top plate
Must be fitted with an O-ring before being mounted in the
Minifors 2 - Benchtop Bioreactor
Accessories
5.4 Blanking Plugs
Blanking plugs are used to seal open ports. There are different
blanking plugs for the different types of port.
Blanking plug, Ø 10 mm
port (see the “Clamping Adaptors and Fastening Screws” section).
Blanking plug, Ø 12 mm
12 mm/Pg13.5 port.
Mounted using a thread.
5.5 Clamping Adapters and Fastening Screws
Clamping adapters are used when mounting the sparger, the various dip tubes and the antifoam/level sensors. The clamping
adapter fixes the component in place and can be used to adjust its
mounting depth.
The clamping adapter must match the outside diameter of the part
being installed and the size of the port in the vessel top plate.
07 February 2019 Page 49 of 214
Fitted with two fixed O-rings (B & C).
head screw is tightened, the component
The fastening screws are used to hold components in place in the
The electrode holder comprises a sheath with a grub screw, a
Minifors 2 - Benchtop Bioreactor
Accessories
Clamping adapter Ø 6/10 mm
When the slotted-head screw is undone, the component with a diameter of 6 mm can be inserted in or removed from the clamping
A
adapter. When the slottedis clamped in the clamping adapter.
B
5.6 Electrode Holder
C
Fastening screw M5
Ø 10 mm ports in the vessel top plate.
Electrode holders are used for mounting sensors (pH, pO2, etc.) in
12 mm/Pg 13.5 ports. They can also be used to adjust the mounting depth of the sensor.
1
guide bar with a fork, and a hollow screw. The wrench for the grub
2
screw is also included in the scope of supply.
1 Sheath
2 Grub screw
3 Guide bar
5
3
4 Fork
5 Hollow screw
4
Page 50 of 214 07 February 2019
Installation depth
17 mm
Hose connection outside diame-
4 mm
INFORMATION
feed needle.
Minifors 2 - Benchtop Bioreactor
Accessories
5.7 Addition Port Adapters and Feed Needles
Addition port adapters and feed needles are used to feed liquid into
the culture vessel. They each come with a tubing connection, are
fitted with a fixed O-ring and are mounted into the four 7.5 mm
ports in the vessel top plate. A single fastening screw is used to
fasten all four addition port adapter(s) and/or feed needle(s) in
place.
Addition port adapter Ø 7.5 mm
Inside diameter 2 mm
Hose connection outside diameter
Addition port adapters protrude as far as the head space of the
vessel, and have very sharp ends with slanted points.
Each culture vessel comes with four addition port adapters as
standard.
4 mm
Feed needle Ø 7.5 mm
Inside diameter 2 mm
ter
Feed needles protrude to below the minimum fill level (= min. working volume) of the culture vessel.
This method of adding liquid allows more precise and regular dosing even when handling small volumes as, unlike the port adapter,
the feed needle does not drip.
The illustration on the left shows only the upper section of the
07 February 2019 Page 51 of 214
The septum collar with inside thread is used to inoculate the cul-
Inside diameter
3.0 mm
Hose connection outside diame-
4.0
INFORMATION
dip tube.
5.8 Septum Collar
5.9 Dip Tubes
Minifors 2 - Benchtop Bioreactor
Accessories
ture in combination with the syringe, injection needle and septum
(inoculation membranes); see the “Inoculation Accessories” section. The septum collar is used to hold the septum in place in the
12 mm/Pg13.5 port.
Dip tubes are open at both ends and are mounted in a vessel top
plate port with a clamping adapter.
Dip tubes are used for a variety of purposes:
For filling the culture vessel after autoclaving. Using a dip tube
prevents foaming.
For adding inoculum.
For sampling. The aseptic Super Safe Sampler system can be
used for sampling.
For harvesting
For siphoning off medium during continuous cultivation
For draining the culture vessel
Depending on the purpose, silicone hoses are connected to the dip
tube via other vessels, sampling systems or, if necessary, hose
networks.
Multiple dip tubes can be used at any one time, providing that
enough vessel top plate ports are available.
Dip tube, straight, Ø 6 mm
ter
The dip tube does not reach as far as the bottom of the vessel.
The illustration on the left shows only the upper section of the
Page 52 of 214 07 February 2019
Fitted with fixed O-ring.
A fastening screw is used to fasten it in the 10 mm vessel top plate
Minifors 2 - Benchtop Bioreactor
Accessories
5.10 Pocket for Temperature Sensor (Pt100)
The pocket is a pipe with a sealed bottom end, and is used to insert the temperature sensor.
Pocket Ø 10 mm
port.
The diagram on the left does not show the fill length of the pocket.
07 February 2019 Page 53 of 214
INFORMATION
system is switched on.
5.11 Exit Gas Cooler
Minifors 2 - Benchtop Bioreactor
Accessories
The basic unit supplies cooling liquid to the exit gas cooler. The
cooling liquid flow rate can be adjusted using the control valve on
the basic unit. The two hoses for water supply (bottom) and return
(top) are connected to the basic unit at the factory and simply connected to the exit gas cooler via the two rapid couplings.
A piece of pressure hose is fitted onto the exit gas cooling pipe and
equipped with an exit gas filter. The hose connections and the exit
gas filter are secured with hose clamps. The hose clamp securing
the exit gas filter is equipped with a screw that can be used to
loosen or tight it manually.
1
2
3
4
a)
b)
5
1 Pressure hose und hose clamp
2 Exit gas filter
3 Hose connections:
a) Water outlet
b) Water inlet
4 Hoses with rapid couplings for water supply and return
(connected before delivery!)
5 Screw thread
The exit gas cooler is fitted with an O-ring before installation.
A screw thread is used to mount it in the 12 mm/Pg13.5 vessel top
plate port.
The exit gas filter must be replaced with a new filter after each
cultivation process.
The exit gas cooler only works when the temperature control
Page 54 of 214 07 February 2019
Two sizes of borosilicate reagent bottle are available for adding re-
INFORMATION
pump heads.
Minifors 2 - Benchtop Bioreactor
Accessories
5.12 Reagent Bottles
1
2
3
agent and feed solution:
250 mL
500 mL
Reagent bottles are fitted before delivery. There are two hose connections on the lid. One is fitted with a short piece of silicone hose
with a filter for pressure equalisation.
The second connection is fitted with a piece of silicone hose at the
other end, inside the bottle.
A 2 m piece of hose is included in the scope of supply for connecting the reagent bottle to the addition port adapter in the culture
vessel and to a pump head.
1 Filter
2 Silicone hose Ø = 2 x 6 mm
3 Cable tie
250 mL reagent bottles are supplied as standard in the unit
package. These fit in the reagent bottle holder that is built into
the vessel holder.
Before delivery with the equipment, the reagent bottles are fitted
with filters and hoses of the correct length, and connected to the
5.13 Sampling System Super Safe Sampler
Basically different systems and also individual components are
available for sampling. This operating manual describes the operation and handling of the aseptic sampling system Super Safe Sampler combined with a dip tube.
The use of the Super Safe Sampler prevents the culture vessel
from contamination when sampling.
07 February 2019 Page 55 of 214
The set consists of a completely pre-assembled group of valves
Minifors 2 - Benchtop Bioreactor
Accessories
Content of the set
with hoses and two syringes. It is connected via silicone hose with
a dip tube.
Valve assembly
5
4
1
1 Sterile filter
2 Check valve
2
3 Luer activated sample valve
4 T-piece
5 Hose
3
The valve assembly consists of a T-piece, two check valves, a
Luer-activated automatic sample valve, a sterile filter, a length of
hose as an adapter for the syringe and another hose for connection to the sample dip tube in the culture vessel.
Page 56 of 214 07 February 2019
The sample valve on the side arm of the T-piece opens by putting
Minifors 2 - Benchtop Bioreactor
Accessories
Principle of function
the Luer connector of the syringe into the valve and closes by removing the syringe. No further handling is necessary.
Unintentional re-introduction of the sample material once it has
been withdrawn is prevented by a check valve. Thus, contamination of the bulk culture is impossible.
Following sampling, a second syringe can be fitted and air pushed
in via the sterile filter, in order to displace culture solution from the
sample hose and the dip tube of the vessel. With a conventional
sampling system, the next sample cannot be taken immediately, as
rinsing of the sampling hose and the immersion tube is necessary.
By previously removing most of the culture in the sampling line,
this sampling system can save culture volume, which is particularly
important with small vessels and/or frequent sampling.
The dead volume of the culture remaining in the group of valves
after flushing with sterile air amounts to a few µl and is negligibly
small. If the withdrawal of a very small sample volume is required,
with minimum possibility of falsification, a small quantity of culture
solution (e.g. 1 ml) can be introduced and rejected before the actual sample is taken.
Designated use
The Super Safe Sampler is designed for aseptic sampling of completely liquid samples.
Solid parts in the sample may lead to clogging of the valves.
Therefore, employing the Super Safe Sampler for solid media is
not recommended.
The Super Safe Sampler is autoclavable (not the syringes!) and for
this reason reusable.
Practical tips for the use of the Super Safe Sampler
Sterility of the culture vessel is ensured at all times without the possible measures mentioned below.
The use of a sterile syringe and sterile caps is only necessary if the
07 February 2019 Page 57 of 214
INFORMATION
Luer lock prevents unwanted movement of the syringe.
The autoclavable pump heads are fitted with PharMed pump hoses
Minifors 2 - Benchtop Bioreactor
Accessories
sample has to be processed under sterile conditions. For sampling,
the same non-sterile syringe can be used repeatedly, without fear
of contamination of the culture vessel.
Aseptic Sampling
For each sample, use a new, sterile syringe with Luer Lock fitting,
in order to ensure the sterility of the sample.
Sterile syringes are consumables and therefore not included in the
set.
The use of another syringe is also possible. But a syringe with
Before fitting the syringe, disinfect the sample valve. Fort this,
spray a commercially available disinfectant onto the valve.
After spraying and after each sampling, close the the sample
valve with a sterile Luer-Lock cap (Dead End Cap) to keep the
valve and sample sterile.
The caps are not included in the kit. Very convenient to use are socalled combi-caps that fit on male and female connectors alike.
Caps that are vented and made of steam sterilisable material can
also be fitted during autoclaving.
5.14 Pump Heads
prior to delivery. Three different hose diameters are available for
different delivery rates:
1.0 mm (standard)
0.5 mm
2.5 mm
For more detailed information about pumps and hoses refer to
main chapter “Technical Data”, chapter “Specification”, “Pumps
Page 58 of 214 07 February 2019
The vessel holder frame has a holding device for four reagent bot-
The reagent bottles are placed in the two holders, and the mount-
Minifors 2 - Benchtop Bioreactor
Accessories
5.15 Vessel Holder with Built-in Holder for Reagent Bottles and Pumps
tles and a holder for the four pump heads.
1 Reagent bottle holder
2 Pump holder
1
2
ing plate with the pump heads is simply pushed onto the pump
holder.
The culture vessel can this be transported and autoclaved as a single unit together with the reagent bottles and pump heads.
5.16 Sterile Filters
Sterile filters are used to protect against contamination in both the
gassing line and the exit gas line. In addition to this, all reagent
bottles used for pressure equalisation must be fitted with an open,
short section of hose with a sterile filter.
07 February 2019 Page 59 of 214
INFORMATION
ideally be replaced after each use.
Retention rate
0.2 µm
Usage
Inlet air (sparger)
Retention rate
0.2 µm
Application
Exit gas
Retention rate
0.3 µm dry
Minifors 2 - Benchtop Bioreactor
Accessories
All the sterile filters in the scope of supply are autoclavable, disposable filters with PTFE diaphragms.
Sterile filters must be clean and dry at all times, and should thus
Ø 37 mm, marked red
Application Inlet air (sparger)
1.5 lit. culture vessels
Ø 50 mm, Type ACro50 TF, no label
Ø 37 mm, marked green
Ø 25 mm, not marked
3.0 and 6.0 L culture vessels
1.0 µm wet
Application Super Safe Sampler
Retention rate 0.2 µm
Diaphragm PTFE
Page 60 of 214 07 February 2019
Application
Reagent bottles (pressure equaliRetention rate
0.45 µm
Diaphragm
PTFE
Hose type
Ø
Application
Pressure hose, fibre-
Water and gas connections
Pressure hose, fibre-
6 x 10 mm
Exit gas filter attachment
Inlet air filter attachment on
Silicone hose
5 x 8 mm
Gassing (sparger)
Pressure hose, trans-
4 x 8 mm
Water supply and return,
Silicone hose, transpar-
2 x 6 mm
Reagent bottles
Attachments
Application
Clamp, 12 mm, INOX
Water and gas connections (on-site)
Hoffmann pinchcock,
Unclamping hose lines, e.g. unused addi-
Hose connectors
Application
Double hose nipple,
Pump heads with hoses to reagent bot-
Minifors 2 - Benchtop Bioreactor
Accessories
5.17 Hoses and Accessories
Ø 25 mm, not marked
sation)
The following hoses and accessories, such as clamps and brackets, are available:
6 x 11.9 mm
glass-woven
(on-site)
glass-woven
Pressure hose, transparent
5 x 10 mm
(on exit gas cooler)
sparger for 3.0 and 6.0 L
TV culture vessels
parent
exit gas cooler
ent
Exit gas filter attachment (on exit gas
cooler)
12 mm, nickel-plated
brass
Cable tie, 2.4 x 86, polyamide
tion port adaptors/feed needles, sparger
hose line, etc.
Hoses for reagent bottles and pumps, inlet air filter, sparger, water supply and return for exit gas cooler, sampling system
dip tube
3/32" x 1/16", PVDF
tles
07 February 2019 Page 61 of 214
Designation
Ø [mm]
Application
O-ring, EPDM
3.53 x 94.84
Top plate gasket, culture
O-ring, EPDM
3.53 x 120.24
Top plate gasket, culture
O-ring, EPDM
2.62 x 10.77
Gasket, port size
Inner gasket for clamping
PTFE ring
120 x 105
Damping ring between
PTFE ring
145 x 130
Damping ring between
PTFE ring
175 x 160
Damping ring between
Accessories for inoculation
Septum (inoculation diaphragm), Ø = 16 mm MVQ silicone, trans-
Tools for screws, grub screws and blanking plugs
Allen key, WAF 2, DIN911
Allen key, WAF 1.27
5.18 O-Rings and Gaskets
Minifors 2 - Benchtop Bioreactor
Accessories
vessel, NW90
vessel, NW115
O-ring, EPDM 3.53 x 148.8
O-ring, EPDM 1.5 x 7.5 Gasket, port size 10 mm
O-ring, EPDM 1.5 x 5.0 Gasket, port size 7 mm
O-ring, EPDM 1.78 x 5.28
Top plate gasket, culture
vessel, NW145
12 mm/Pg13.5
adaptor for 10 mm ports
Flat gasket, silicone 32 x 42 x 2
5.19 Inoculation Accessories and Tools
The following inoculation accessories and tools are used:
parent, for 12 mm/Pg13.5 ports
Sterile disposable syringe, Luer, 10 mL, inside Ø 14.35 mm
Sterile hollow needle, 20G, L = 40 mm/Ø = 0.9 mm
glass vessel and vessel
holder, NW90
glass vessel and vessel
holder, NW115
glass vessel and vessel
holder, NW145
Gasket for reagent bottle
lid (all sizes)
For grub screws on impellers
For grub screws
Page 62 of 214 07 February 2019
Hexagon socket spanner, WAF 17
Minifors 2 - Benchtop Bioreactor
Accessories
5.20 Starter Kit
5.21 Service Sets
For blanking plugs in 12 mm/Pg13.5 ports
Torx wrench, TX25
For screws on the thermal block adapter
Each equipment package comes with a starter kit with a variety of
hoses, attachments, inoculation accessories and tools. A detailed
contents list is included in each starter kit.
5.22 Auxiliary Supplies
Service sets with O-rings, gaskets, sterile filters, hoses, etc. to fit
each vessel size are available separately. A detailed contents list is
included in the service set.
The term “auxiliary supplies” covers all the substances and materials required for operation and/or maintenance that cannot be considered part of the equipment or the system.
pH Buffers
pH buffers are used to calibrate the pH sensors. 250 mL bags are
available for the following buffers:
pH 4.04
pH 7.01
07 February 2019 Page 63 of 214
WARNING
vere property damage.
WARNING
be carried by one person alone.
6 Transport and Storage
The following specifications are based on transport and storage of
an unpacked equipment at the provider’s site.
6.1 Transport
Improper transport, the use of incorrect auxiliary equipment and
careless handling of the equipment may lead to injuries and se-
The following points must be observed when transporting the
equipment internally (relocation):
Always work in pairs and use suitable auxiliary equipment
The entire equipment (basic unit and culture vessel) contains
Especially when using auxiliary tools, it is important to observe
Minifors 2 - Benchtop Bioreactor
Transport and Storage
when transporting the equipment.
delicate glass parts.
that the equipment’s centre of gravity is not in the middle.
The entire equipment (basic unit and culture vessel) is too
heavy to be carried by one person alone.
Even the basic unit on its own exceeds the weight that should
Page 64 of 214 07 February 2019
Minifors 2 - Benchtop Bioreactor
Transport and Storage
6.2 Storage
Before each time they are put into storage, decontaminate,
thoroughly clean and dry the culture vessel and all accesso-
1
ries
.
Store the equipment and its components clean, dry and pro-
tected against dust, dirt and liquids.
Store the equipment and its components in a cool place with
low air humidity but protected against frost.
Storage temperature: 5°C – 55°C
Relative air humidity, non-condensing: 10% - 95%.
Protect the equipment from aggressive media, direct sunlight
and mechanical vibrations.
1)
Maintain and store sensors produced by other manufacturers in accord-
ance with the separate documentation.
07 February 2019 Page 65 of 214
WARNING
erating manual precisely.
Installation and Initial Operation
7 Installation and Initial Operation
To set up and connect the equipment, note the following:
7.1 General Location Requirements for Installation
Faulty installation may lead to dangerous situations or severe
loss of property.
Follow the installation and commissioning instructions in this op-
Minifors 2 - Benchtop Bioreactor
The following requirements must be met for the installation of the
equipment:
The figures and ranges specified in the chapters “Technical
Data, Connection Values” and “Technical Data, Operating
Conditions” must be observed.
The equipment must only be installed inside a laboratory or a
laboratory-like environment.
The installation site must be level, sufficiently stable and able
to bear loads.
There must not be any sources of electrical interference near
the equipment.
7.2 Minimum Distances
To operate and maintain the unit it must be installed with a minimum spacing of 150 mm from walls, ceilings or other equipment.
7.3 Connecting the Equipment to On-Site Supply Lines
The following chapters describe which connection requirements
must be fulfilled on site and how the equipment is connected to onsite supply lines.
Page 66 of 214 07 February 2019
ATTENTION
age or failure of the equipment.
Minifors 2 - Benchtop Bioreactor
Installation and Initial Operation
7.3.1 Power Supply
Connection conditions
The in-house power supply to the equipment must meet the following requirements:
Single-phase, constant power supply
120/230 VAC 50/60 Hz
Connection
To connect the basic unit to the in-house power supply, proceed as
follows:
Procedure 1. Insert the power cable supplied into the connector socket on
the equipment.
2. Insert the cable into the in-house power supply.
7.3.2 Water Supply and Return
Connection conditions
The in-house water supply to the unit, as well as the drainage of
the water, must meet the following requirements:
“Very soft” or “soft” water quality (CaCO
-1
L
to 1.5 mmol L-1)
Not observing the water quality requirements may lead to dam-
Constant water supply at a pressure of 2 ± 1 bar
Manometer to check the primary pressure available
The drain is heat-resistant and without back pressure
Connection
To connect the basic unit to the in-house water supply and drainage, proceed as follows:
Procedure 1. Cut the required quantity of the supplied pressure hose (Ø = 6
x 11.9 mm).
concentration 0 mmol
3
07 February 2019 Page 67 of 214
ATTENTION
Only use dry, clean and oil-free gases.
7.3.3 Gas Supply
Minifors 2 - Benchtop Bioreactor
Installation and Initial Operation
2. Position the pressure hoses on the appropriately marked hose
nozzles on the basic unit.
3. Connect the hoses to the in-house water supply and drainage.
4. Secure the hoses with hose clamps to prevent slipping.
5. Check to ensure that the hoses neither have kinks nor are
able to kink and that connections and hoses do not have any
leaks.
Connection conditions
The in-house gas supply to the equipment must meet the following
requirements:
Constant gas supply at a pressure of 2 ± 0.5 bar
Gas(es) is/are dry, clean and free of oil and dust
Recommended compressed-air quality as per DIN ISO 8573-
1: Class 1,2,3,4
The use of impure gases can lead to blockage of the sterile filter
and damage the mass flow controller.
Connection
To connect the basic unit to the in-house gas supply, proceed as
follows:
Procedure 1. Cut the required quantity of the supplied pressure hose (Ø = 6
x 11.9 mm).
Only use hoses supplied by the manufacturer.
2. Position the pressure hoses on the appropriately marked hose
nozzles on the basic unit.
3. Connect the hoses to the in-house gas supply.
4. Secure hoses with hose clamps to prevent slipping.
5. Check to ensure that hoses neither have kinks nor are able to
kink and that connections and hoses do not have any leaks.
Page 68 of 214 07 February 2019
WARNING
when the equipment is not in use.
ATTENTION
circuit that could damage the control electronics.
Minifors 2 - Benchtop Bioreactor
Installation and Initial Operation
7.3.4 Exit Gas
The use of inappropriate or damaged hoses and/or inappropriate fixing may lead to leakage of gases. Depending on the gas
in question, there may be a danger of gas explosion and/or danger of suffocation as well as a hazard for the health of the operator.
Always close the gas supply before a hose is removed and
On site, it must be ensured that:
the exit gas is dissipated securely by means of a suitable,
gas-tight hose.
the working environment and/or the laboratory/laboratory-like
facility is equipped with a sufficient ventilation system, depending on the application.
7.4 Connecting the Motor Cable
The motor is controlled directly via the basic unit and is connected
to it via the motor cable.
For routine operation, it is not necessary to plug in and unplug the
motor cable. The connected motor is only coupled before cultivation. For details, see the main chapter “Before Cultivation” and the
“Connecting the Motor” chapter.
If the motor cable is connected to or disconnected from the motor while the equipment is switched on, there is a risk of a short
To connect the motor cable, proceed as follows:
Procedure 1. Insert the (angled) plug of the motor cable into the socket on
the rear of the basic unit.
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7.5 Test Run
Minifors 2 - Benchtop Bioreactor
Installation and Initial Operation
2. Insert the other plug into the socket on the motor.
Depending on the size of the culture vessel, the large (lefthand) or small (right-hand) motor is supplied.
In order to become familiar with the basic functions of the equipment before the first cultivation, a short test run can be executed.
The test run comprises:
Temperature control (cooling / heating)
Stirring
Gassing
Compressed air of the stated quality (see chapter “Gas Supply”) is
used for gassing.
To avoid calcium deposits, demineralised water is recommended
for filling the vessel.
The following description of the test run does not detail handling of
individual components, e.g. exit gas cooler, stirrer, sparger etc. Detailed descriptions of their handling are given in the corresponding
chapters of the main chapter “Before Cultivation”.
For details on operation, see the main chapter “Operation”.
7.5.1 Preparation Test Run
Before starting the test run, check and ensure the following:
the equipment is correctly connected to the water, power and
gas supply and is operational
the motor cable is connected to the basic unit and the motor.
The following work is to be executed before the test run:
Procedure 1. Remove the vessel top plate and put it aside carefully.
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ATTENTION
of components.
Minifors 2 - Benchtop Bioreactor
Installation and Initial Operation
If the vessel top plate presses against long components such as
the stirrer shaft etc., they could bend because of the weight of
the top plate.
Always position the vessel top plate so that it does not lie on top
2. Fill the culture vessel with water – preferably demineralised –
to the working level.
3. Ensure that the stirrer and sparger are mounted; if necessary,
mount them.
4. Fit the top plate and secure it.
5. Screw the exit gas cooler into the port on the vessel top plate
port.
The exit gas cooler is equipped with a new exit gas filter
in the factory.
6. Connect the exit gas cooler to pre-fitted hoses on the basic
unit; to this end, follow the symbols on the basic unit:
water inlet on bottom of exit gas cooler / water outlet at
top of exit gas cooler.
7. Close all remaining open ports with blanking plugs.
8. Hang the culture vessel on the basic unit.
9. Connect the sparger to the gassing system (compressed air)
on the equipment; to this end, attach the gassing hose to the
nozzle on the inlet air filter.
The sparger is equipped with hose and the inlet air filter
in the factory.
The gassing hose is attached to the basic unit in the factory.
10. Insert the temperature sensor as far as it will go into the
pocket in the top plate.
11. Couple the motor.
12. Switch on the equipment at the main switch and wait until it
has started up.
7.5.2 Cooling System
To activate the cooling system, proceed as follows:
Procedure 1. On the operating panel, set a low setpoint for the Temperature
parameter, e.g. 10 °C, in order to activate the water supply to
the temperature control system.
07 February 2019 Page 71 of 214
CAUTION
block adapter are touched!
Minifors 2 - Benchtop Bioreactor
Installation and Initial Operation
2. Start the Batch (process) using Start Batch and switch on the
Temperature parameter.
3. All parameters except for Temperature remain switched off;
switch them off if necessary.
You should now hear water flowing into the temperature control system.
The water supply to the exit gas cooler should be activated,
too now.
4. Use your hands to check whether the exit gas cooler and thermal block and/or adapter are beginning to cool down.
As soon as the temperature control circuit is full, water will
flow out of the water outlet (H2O OUT) of the basic unit.
For the rest of the procedure, allow the Batch to run with the temperature switched on.
7.5.3 Stirring
Batch (process) is running with temperature switched on
To test the stirrer, proceed as follows:
Procedure 1. On the operating panel for the Stirrer parameter, set a low set-
-1
point, e.g. 200 min
.
2. Switch on the Stirrer parameter.
For the rest of the procedure, allow the Batch to run with the temperature switched on and the stirrer running.
7.5.4 Heating and Adjusting Temperature
Batch (process) is running with temperature switched on and
stirrer running
To test the heating and adjust the temperature, proceed as follows:
Procedure 1. On the operating panel, set a high setpoint for the Tempera-
ture parameter, e.g. 45 °C.
The water supply for cooling is stopped; the system heats up.
Risk of minor burns if the heated thermal block and thermal
Page 72 of 214 07 February 2019
INFORMATION
and the correct setting in the Vessel Type menu.
CAUTION
tion, is touched!
Minifors 2 - Benchtop Bioreactor
Installation and Initial Operation
2. Wait until the temperature has adjusted to the setpoint.
For the rest of the procedure, allow the Batch to run with the temperature switched on and the stirrer running.
7.5.5 Gassing
Batch (process) is running with temperature switched on and
stirrer running
To test the gassing, proceed as follows:
Procedure 1. On the operating panel for the AirFlow parameter, set a low
setpoint, e.g. 1.0 L min
2. Switch on the AirFlow parameter.
-1
.
The maximum airflow depends on the size of the culture vessel
3. Ensure that all other gas parameters (Gas2Flow, TotalFlow,
GasMix) are switched off.
If the gassing is working, air bubbles now form in the water in
the culture vessel.
7.5.6 End of Test
After all parameter setpoints have been reached, the test can end
here. The inoculation that now takes place during normal operation
is not relevant to the test run.
Proceed as follows:
Procedure 1. On the operating panel, press Inoculate and then Stop Batch
to stop the Batch (process).
2. Switch the equipment off at the main switch.
3. Shut off the supply lines.
4. Let the motor cool down.
Risk of minor burns if the motor, which heats up during opera-
07 February 2019 Page 73 of 214
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Installation and Initial Operation
When the motor has cooled down:
5. Uncouple the motor from the vessel and place it on a clean
and dry work surface.
6. Empty the culture vessel.
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INFORMATION
Do not use silicone grease; this can affect sterilisation results.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
8 Before Cultivation
The following chapters describe all the preparatory work before
starting the cultivation process. This essentially comprises:
Preparing and autoclaving the culture vessel:
Checking the gaskets (O-rings) on component parts and
culture vessel
Mounting component parts
Filling or moistening the culture vessel
Preparing sensors and other accessories
Autoclaving
Connecting the culture vessel and preparing for cultivation:
Hanging the culture vessel into place on the basic unit
and connecting cables and hoses between the culture
vessel and the equipment
Filling the vessel if necessary
Preparing sensors and other accessories
8.1 Preparing and Autoclaving the Culture Vessel
All accessories required for later cultivation must be prepared and
mounted accordingly and autoclaved together with the culture vessel.
Certain accessories are already mounted when the equpment is
delivered.
8.1.1 Checking Gaskets (O-Rings)
O-rings are used to seal all openings on the vessel and top plate.
The top plate and all internal fittings or connections are thus
equipped with O-rings. Before every use, the O-rings must be
checked that they are present, undamaged and correctly seated.
Damaged O-rings must be replaced.
Wet the O-rings with 70% alcohol or a little water to facilitate removing and replacing O-rings or component parts with O-rings.
Carry out this check as follows:
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Procedure
INFORMATION
Septum collars are sealed with a septum. No O-ring is used!
INFORMATION
the same height as the surface of the medium.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
1. Check the O-ring for sealing the top plate for damage and to
ensure that it is positioned correctly in the groove on the vessel flange.
2. Ensure that every component part is equipped with an intact
O-ring:
Check that the O-rings are correctly positioned and are undamaged. If necessary, reposition or replace. If component
parts are fitted into other component parts (clamping adaptor), there must also be an O-ring between them.
8.1.2 Mounting the Impellers
Procedure
To mount the impellers to the stirrer shaft, proceed as follows:
1. Slide the impeller onto the stirrer shaft.
2. Set the desired height.
3. Tighten the grub screws on the impeller with the Allen key.
To avoid unnecessary foam formation, do not fit the impeller at
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Before Cultivation
8.1.3 Mounting Dip Tubes and Spargers
Straight spargers and dip tubes can be mounted to the outside of
the vessel top plate. Curved spargers and dip tubes can only be
mounted to the inside of the vessel top plate.
Since this unit uses curved spargers and straight dip tubes, mounting to the inside of the vessel top plate is described here. This
means that the vessel top plate is still removed.
During mounting, ensure that the sparger or the dip tube does not
come into contact with other component parts (stirrer). The sparger
is positioned below the stirrer shaft.
Proceed as follows:
Procedure 1. Ensure that the clamping adapter is equipped with an interior
or exterior O-ring; if necessary, attach O-ring(s).
2. Insert the clamping adapter into the intended port and fix it
with a fastening screw.
3. Loosen the slotted-head screw at the clamping adapter.
4. Insert the sparger/dip tube into the clamping adapter from be-
low.
07 February 2019 Page 77 of 214
Procedure
the two opposing recesses on the flange fit with the bolt on the
Minifors 2 - Benchtop Bioreactor
Before Cultivation
5. Set the desired mounting depth.
6. Tighten the slotted-head screw.
8.1.4 Inserting the Vessel into the Vessel Holder
To insert the glass vessel into the vessel holder, proceed as follows:
1. Place the flange onto the ring of the vessel holder:
ring.
2. Place the damping ring onto the flange.
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Before Cultivation
3. Insert the vessel carefully.
8.1.5 Inserting the Baffles
Procedure
Proceed as follows:
1. Insert the baffles carefully into the glass vessel.
07 February 2019 Page 79 of 214
INFORMATION
water per litre of total volume).
Procedure
8.1.6 Moistening/Filling the Culture Vessel
If the medium in the culture vessel is to be autoclaved, this can
take place before the top plate is put in position and the additional
component parts are mounted.
Note the following about filling the culture vessel before autoclaving:
Before autoclaving, only top up with heat-resistant media.
During autoclaving, evaporation may result in a loss of volume
and thus to increased salt concentration in the medium. If necessary, top up with sterile water.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
Development of steam is not possible when autoclaving an
empty and dry culture vessel. Successful sterilisation is not
guaranteed.
Ensure that there is liquid in the culture vessel (approx. 10 mL of
8.1.7 Fitting the Vessel Top Plate
Proceed as follows to fit and fix the vessel top plate:
1. Fit the O-ring for the top plate gasket into the groove on the
2. Place the top plate carefully and with the correct alignment
edge of the vessel.
into position and ensure that component parts do not touch
the baffles.
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ATTENTION
specify that they must be tightened by hand
Procedure
Minifors 2 - Benchtop Bioreactor
Before Cultivation
3. Tighten the knurled screws on the top plate by hand (no tool!)
crossways.
If the screws are tightened too much, components may be damaged, which can result in failure of the equipment. The screws
may not be tightened with a tool under any circumstances.
This applies to all screw connections where the instructions
8.1.8 Mounting the Blanking Plugs
For mounting the different blanking plugs, proceed as follows:
Ø 10 mm ports
1. Insert the blanking plug with fixed O-ring into all unused ports.
2. Fix with a fastening screw.
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Procedure
Procedure
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Before Cultivation
Ø 12 mm / Pg13.5 ports
1. Insert the O-ring and blanking plug into all unused ports.
2. Tighten by hand.
3. Use the hexagon socket spanner to make it hand-tight.
8.1.9 Mounting Addition Port Adapters
Proceed as follows:
1. Insert the addition port adapters with a fixed O-ring into the
four 7.5 mm ports.
2. Fix it with the fastening screw.
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Procedure
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Before Cultivation
8.1.10 Mounting the Feed Needle(s)
The procedure for mounting one or more feed needle(s) instead of
addition port adapters is the same as for the mounting of the addition port adapters. For details, see the chapter “Mounting Addition
Port Adapters”.
8.1.11 Mounting the Pocket for Temperature Sensor (Pt100)
Proceed as follows:
1. Insert the pocket with the fixed O-ring into the 10 mm port.
2. Fix it with the fastening screw.
8.1.12 Equipping the Port with a Septum Collar and Septum for Inoculation
Procedure 1. Ensure that there is no O-ring in the port; if there is, remove it.
For later inoculation with a syringe, the 12 mm/Pg13.5 port in the
top plate must be prepared as follows:
07 February 2019 Page 83 of 214
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Before Cultivation
2. Insert the septum (inoculation diaphragm) into the port.
3. Screw the septum collar into the port by hand.
4. Insert the blanking plug equipped with an O-ring into the sep-
tum collar and screw it tight by hand.
If necessary, use the hexagon socket spanner to make it
hand-tight.
8.1.13 Preparing the Dip Tube/Addition Port Adapter for Inoculation
If later inoculation is to be carried out by means of a dip tube or
addition port adapter, proceed as follows:
Procedure 1. Fit the dip tube with the clamping adapter or addition port
adapter in the port.
2. Place a piece of silicon hose onto the dip tube/addition port
adapter.
Page 84 of 214 07 February 2019
INFORMATION
formation is expected.
CAUTION
uum in the culture vessel may occur during autoclaving.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
3. Equip the hose for a sterile hose connection. (Depending on
the application: rapid coupling, sterile connector or weldable
hose with sterile filter).
4. Secure the hose transition points with cable ties.
8.1.14 Mounting the Exit Gas Cooler
Mount the exit gas cooler as follows:
Procedure 1. Attach the O-ring to the thread of the exit gas cooler.
2. Screw the exit gas cooler onto the thread in the
12 mm/Pg13.5 port by hand.
3. Align the exit gas cooler to ensure that handling of other com-
ponent parts is impaired as little as possible.
4. Check to ensure that the exit gas filter is fitted securely.
5. Cap the exit gas filter loosely with a little aluminium foil
A humidifier bottle with antifoam reagent can be installed between exit gas cooler and the exit gas filter if significant foam
Take the following into account for autoclaving:
Only use a new, clean and dry exit gas filter and fix it in such a
way that it cannot slip.
ALWAYS keep the exit gas line - hose at the exit gas cooler
with secured exit gas filter - open.
If pressure equalisation does not take place via a top plate
opening or the mounted exit gas cooler, overpressure or vac-
07 February 2019 Page 85 of 214
ATTENTION
foil during autoclaving!
8.1.15 Preparing the Sensors
Minifors 2 - Benchtop Bioreactor
Before Cultivation
All sensors that come into contact with the medium are mounted
before autoclaving and are sterilised together with the culture vessel.
Note the following about all sensors:
Mount all sensors by hand – do not use any tools!
Mount the sensors in such a way that they cannot come in
contact with other components or the glass vessel.
If the mounting depth of is adjustable (mounting with electrode
holder/clamping adaptor), make sure the mounting depth is
set correctly prior to autoclaving, as later adjustment represents a contamination risk.
pH sensor
Calibrate the pH sensor on the operating panel before mount-
ing and autoclaving.
sensor
pO
2
Mount the pO
to the flow and there is no risk of bubbles collecting. (Calibration is carried out AFTER autoclaving)
pH sensor & pO
For vessels with a nominal width of 90 and 145: screw the
sensors directly into 12 mm/Pg13.5 port.
For vessels with a nominal width of 115: mount the sensors
with an electrode holder.
sensor in such a way that it has good access
2
sensor
2
Risk of damage to the pH and pO2 sensors. Covering the sensor
heads with aluminium foil during autoclaving may lead to water
gathering under the film, thus damage the contacts on the sensor head.
pH and pO
sensor heads may NOT be covered with aluminium
2
For details on the safety, technical data, usage and maintenance
requirements for the pH and pO
sensors, see the separate docu-
2
mentation provided by the sensor manufacturers.
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INFORMATION
using the operating panel is no longer necessary.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
8.1.16 Calibrating the pH Sensor
For a reliable pH measurement, a 2-point calibration with an upper
and lower reference buffer must be carried out before each cultivation. The pH sensor must be calibrated before autoclaving. This is
carried out on the operating panel.
Procedure 1. Connect the sensor cable. (For more details, see the chapter
“Connecting the pH Sensor”).
2. Switch on the equipment using the main switch.
The operating panel is switched on automatically and the system is started.
3. Calibrate the pH sensor in accordance with the detailed de-
scription in the main chapter “Operation”, chapter “Calibrating
the pH Sensor”.
If the pH sensor has already been calibrated before connection
to the system, the bioreactor will use this data and calibration
8.1.17 Mounting a Sensor into a 12 mm Port
For culture vessels with nominal widths of 90 and 145, sensors can
be screwed directly into 12 mm/Pg13.5 ports. To do so, proceed as
follows:
Procedure 1. Slide the O-ring onto the sensor.
2. Insert the sensor into the port.
3. Screw the sensor on its thread into the port by hand.
07 February 2019 Page 87 of 214
8.1.18 Mounting Sensors with Electrode Holder
For the mounting of a sensor in a 12 mm/Pg13.5 port for culture
vessels with a nominal width of 115, an electrode holder must be
used.
To do so, proceed as follows:
Procedure
1. On the electrode holder, lightly loosen the grub screw in the
support guide with the key.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
2. Pull the support guide from the guide bar.
3. Insert the sensor into the support guide and tighten it.
Page 88 of 214 07 February 2019
thread pointing
Minifors 2 - Benchtop Bioreactor
Before Cultivation
4. Insert the sensor into the hollow screw with the
in the downward direction.
5. Fit the fork of the guide bar into the groove of the hollow
screw.
6. Push the hollow screw and the guide bar together upwards
and insert the guide bar into the hole of the support guide.
7. Slide the O-ring onto the sensor and insert the sensor into the
port.
8. Adjust the sensor to the desired height.
07 February 2019 Page 89 of 214
Screw the sensor on the hollow screw into the port and tighten
Minifors 2 - Benchtop Bioreactor
Before Cultivation
9.
it.
10. Tighten the grub screw in the support guide with the key.
8.1.19 Mounting the Antifoam Sensor
Please note the following points for mounting:
The antifoam sensor is equipped with transparent insulation
that must be intact, as otherwise a continuous signal
“Foam/liquid detected” may be generated.
The sensor head must not touch the clamping adaptor, other-
wise a continuous short-circuit is generated, indicating
“Foam/liquid detected”.
The clamping adapter on the sensor must be equipped with
Procedure 1. Remove the protective cap from the sensor.
an intact O-ring.
Proceed as follows for mounting:
2. Insert the sensor into the port.
Page 90 of 214 07 February 2019
ATTENTION
damaged.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
3. Fix the clamping adapter with the fastening screw.
4. Loosen the slotted-head screw at the clamping adaptor.
5. Set the desired mounting depth of the sensor carefully.
6. Tighten the slotted-head screw carefully.
If the sensor is fixed too tightly in the clamping adapter, or the
mounting depth of the sensor is changed while the screw on the
clamping adapter is tightened, the sensor insulation may be
07 February 2019 Page 91 of 214
INFORMATION
sion.
Procedure
8.1.20 Preparing the Super Safe Sampler
The following figures are for general purposes of comprehen-
In order to prepare the Super Safe Sampler sampling system for
autoclaving, proceed as follows:
1. Attach the hose of the valve group on the dip tube.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
2. Secure the hose with a cable tie.
3. Tighten the sample valve carefully by hand in a clockwise di-
rection.
This ensures that the non-return valve/sample valve screw
connection is tight.
Page 92 of 214 07 February 2019
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Before Cultivation
4. Turn the sterile filter carefully by hand in a clockwise direction.
This ensures that the non-return valve/sterile filter screw connection is tight.
5. Cover the valve group loosely with aluminium foil.
6. Clamp off the hose on the dip tube.
8.1.21 Mounting the Sparger Hose and the Inlet Air Filter
The sparger must be equipped with the hose and inlet air filter before autoclaving.
To do so, proceed as follows:
Procedure 1. Cut a short piece of hose:
1.5 L culture vessels: silicone hose Ø = 2 x 6 mm
3.0 L and 6.0 L culture vessels: pressure hose, transpar-
ent, Ø = 5 x 10 mm.
2. Fit the inlet air filter to the hose piece:
1.5 L culture vessel: filter with red marking, Ø = 37 mm, fit it in
the direction of the air flow to the hose end. The nozzle with
the red INLET marking remains exposed.
07 February 2019 Page 93 of 214
ATTENTION
Minifors 2 - Benchtop Bioreactor
Before Cultivation
3.0 L and 6.0 L culture vessels: filter without marking, Ø =
50 mm, fit it in either direction to the hose end.
3. Fit the hose to the sparger.
The figure to the left shows an inlet air filter for 1.5 L culture
vessels as an example.
4. Secure the ends of the hose with the cable tie.
5. Clamp off the hose with a hose clamp.
6. Lightly cap the inlet air filter with aluminium foil.
8.1.22 Preparing the Reagent Bottles, Pumps and Hoses
250 mL reagent bottles are supplied as standard in the equipment
package. These fit in the reagent bottle holder that is built into the
vessel holder. Before delivery with the equipment, the reagent bottles are equipped with filters for pressure equalisation and hoses of
the correct length, and connected to the pump heads.
Damaged hoses and/or clogged sterile filter may lead to undesired pressure conditions in the reagent bottles.
– Ensure each reagent bottle is equipped with an open pres-
sure equalisation line with a clean and dry filter.
– Only use clean, intact hoses and they are firmly attached.
Page 94 of 214 07 February 2019
INFORMATION
culture vessel do not have any tensions or kinks.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
Below is a detailed description of how reagent bottles are equipped
properly and connected to the pumps and culture vessel.
Connecting the reagent bottles to the pumps and culture vessel
To connect the reagent bottles with the pumps and the culture vessel, proceed as follows:
Procedure 1. Cut two long silicone hoses (Ø = 2 x 6 mm) per pump/reagent
bottle.
The length of the silicone hoses must be selected to ensure that
the hose connections between the reagent bottles, pumps and
2. Thoroughly rinse the silicone hoses with distilled water.
3. Connect the silicone hoses and pump hoses of the pump
heads with hose connectors.
For the Fill function:
Right-hand side = suction side = hose line to reagent
bottle.
Left-hand side = pumping side = hose line to culture
vessel.
See arrow for direction of rotation.
4. Secure with cable ties.
Connection between pumps and culture vessel
Procedure
1. Fit silicone hoses for base, acid and feed to addition port
adapter and/or feed needle(s) and secure them with cable
ties.
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ATTENTION
Use only non-corrosive acids, e.g. phosphoric acid, instead.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
2. Attach the silicone hose of the antifoam pump to the mounted
antifoam sensor in the culture vessel and secure it with a cable tie.
Connection between reagent bottles and pumps
Procedure 1. Ensure that a hose is fitted inside the reagent bottles at the
exposed hose connection (without sterile filter); fit one if not:
a) the end of the hose does not touch the bottom of the bot-
tle, otherwise the hose may get sucked against the bottom and no longer be able to pump liquid.
b) the end of the hose is cut diagonally. In this case the
hose end can touch the bottom of the bottle.
2. Label the reagent bottles in accordance with their content.
3. Depending on the application: Fill the reagent bottles with rea-
gents and reclose them with their lid.
Usage of the highly corrosive hydrochloric acid HCl as reagent
leads to damage to components made of stainless steel such as
e.g. component parts or the top plate.
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INFORMATION
the reagent bottle after sterilising.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
Fill reagent bottles with heat-resistant reagents only. Sterilise
non-heat-resistant feed solution separately and only transfer it to
4. Place the reagent bottles in reagent bottle and pump holders.
5. Attach the correct silicone hoses to available hose connec-
tions of each reagent bottle and secure them with cable ties.
6. Close silicone hoses with clamps as close as possible to the
7. Ensure that:
8. Cap the filter loosely with aluminium foil.
8.1.23 Sterile Hose Connections
If additional vessels are needed and these can only be connected
to the culture vessel after autoclaving, such as vessels for the inoculum or bottles for sampling etc., rapid couplings (male/female),
sterile connectors or – if weldable hoses are used – a hose welding
device can be used to form a sterile connection.
The connection pieces must be fitted to the appropriate hoses before autoclaving. Rapid couplings are connected after autoclaving
in a sterile workbench. Sterile connectors and hose welding devices allow sterile connecting without a sterile workbench.
hose connections of the reagent bottles to ensure that no reagent can flow into the culture vessel.
each reagent bottle is connected with the appropriate
pump according to its contents. (Base to base pump,
etc.)
filters are clean and dry; short hose line is open.
07 February 2019 Page 97 of 214
8.1.24 Setting the Pumps
If the pumps are not used with the default settings, we recommend
that the appropriate settings are now made on the operating panel.
It is possible, for example, to estimate and display the volume (in
mL) that has been pumped since the Batch (process) started. To
this end, the diameter of the hose used must be selected.
For details on the pumps and the setting options, see the main
chapter “Operation”, the “PUMPS parameter group” chapter and
the associated sub-chapters.
8.1.25 Removing the Pump Heads
To remove the pump heads from the basic unit, proceed as follows:
Procedure 1. Swing open the pump cover.
Minifors 2 - Benchtop Bioreactor
Before Cultivation
2. Remove the mounting plate with the pump heads from the
drive shafts by holding the two handles.
3. Place the mounting plate with the pump heads onto the pump
holder on the vessel holder.
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ATTENTION
the drive hub!
Culture vessel
Drive hub is equipped with cone plug.
There is liquid in the culture vessel (autoclavable medium or approx. 10 mL water per litre working volume).
Reagent bottles, hoses and pumps
Reagent bottles are exclusively filled with autoclavable reagents, correctly labelled and connected with the
Reagent bottles are equipped with filters for pressure equalisation
Minifors 2 - Benchtop Bioreactor
Before Cultivation
8.1.26 Fitting the Cone Plug for Drive Hub
In order to prevent the penetration of condensation water into the
dive hub during autoclaving, the cone plug provided in the starter
set must be fitted.
Risk of loss of property du penetration of condensation water
into the drive hub!
Always autoclave the culture vessel with the cone plug fitted to
Proceed as follows:
Procedure
1. Plug the cone plug into the opening of the drive hub.
8.1.27 Checklist Before Autoclaving
Check and ensure the following items before autoclaving:
All necessary O-rings are fitted.
All unused ports are closed with blanking plugs
Connection for inoculation is equipped with septum, septum collar and blanking plug
culture vessel and the pump heads via hoses.
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