OLS CERO Manual

CELL CULTURE SIMPLIFIED |

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Important Notice
This Operators Guide may not be used or reproduced in any way whatsoever without the
express written consent of OMNI Life Science GmbH & Co. KG
Copyright © 2018 OMNI Life Science GmbH & Co. KG (OLS), All Rights Reserved.
Contents of this Operators Guide are subject to change without prior notice. Technical
modifications reserved. Greatest possible care was used on the correctness of the information
in this Operators Guide. If errors should be discovered nevertheless, OLS is pleased to be
informed about it. Regardless of this, OLS cannot assume any liability for errors in this
Operators Guide or for their consequences.

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Preface –Welcome to the World of Cell Culture Simplified
Congratulations on your purchase of CERO!
The CERO is a versatile, walk-away bench-top instrument capable of handling four independent
and high density micro carrier or suspension cell cultures. In addition, the CERO eliminates
traditional peripheral cell culture instruments, such as incubators and centrifuges, and
minimizes manual handling.
With proper care and maintenance, your new CERO system will serve you faithfully. To learn
about the proper care and maintenance of your investment, please take the time to read this
Operators Guide.
We at OLS thank you for purchasing this product.
Welcome to the world of automated microcarrier and suspension cell culture!

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TABLE OF CONTENTS
TABLE OF CONTENTS............................................................................................5
PROLOGUE ...........................................................................................................9
I. Revision History ................................................................................................................. 9
II. Contact Addresses............................................................................................................. 9
III. Declaration of Conformity ...............................................................................................10
IV. Warranty.........................................................................................................................10
1. GENERAL INFORMATION ................................................................................11
1.1 Intended Use ..................................................................................................................11
1.2 About this Operators Guide............................................................................................11
1.3 Operation .......................................................................................................................11
2. SAFETY PRECAUTIONS AND HAZARDS ............................................................12
2.1 General Precautions .......................................................................................................12
2.1.1 Instrument.................................................................................................................... 12
2.1.2 Operating the CERO...................................................................................................... 12
2.2 Hazards of CO2................................................................................................................13
2.3 Electrical Safety Precautions...........................................................................................13
2.4 Biohazard Precautions....................................................................................................13
3. PRODUCT DESCRIPTION .................................................................................14
3.1 Culturing Cells in the CERO .............................................................................................14
3.1.1 Parameters, Protocol and Experiment ......................................................................... 14
3.1.2 Inoculations Protocol.................................................................................................... 15
3.1.3 Culture Protocol ........................................................................................................... 16
3.1.4 Harvesting Protocol ...................................................................................................... 16
3.2 Allgemeine Beschreibung des CERO ...............................................................................17
3.3 Culture Chamber ............................................................................................................17
3.3.1 Sealed Culture Chamber............................................................................................... 17
3.3.2 CO2and Temperature Control...................................................................................... 18
3.3.3 Control of the Cell Suspension ..................................................................................... 18
3.4 User Interface.................................................................................................................19
3.5 CERO Software ...............................................................................................................20
3.5.1 Main Menu ................................................................................................................... 21
3.5.2 Experiments Menu ....................................................................................................... 21

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3.5.3 System Menu ................................................................................................................21
3.5.4 Outputs Menu ...............................................................................................................21
3.5.5 Run Menu......................................................................................................................21
3.6 Electronics ..................................................................................................................... 22
3.6.1 Mains Power Socket......................................................................................................22
3.6.2 Fuse Holder ...................................................................................................................22
3.6.3 Power On / Off Switch...................................................................................................22
3.6.4 Open Cover Button .......................................................................................................23
3.6.5 Communications Ports ..................................................................................................23
4 INSTALLATION ................................................................................................. 24
4.1 Product Content............................................................................................................. 24
4.2 Requirements ................................................................................................................ 24
4.3 Installation Pocedure ..................................................................................................... 25
5. OPERATION .................................................................................................... 27
5.1 Switch ON the CERO....................................................................................................... 27
5.2 Set Temperature ............................................................................................................ 27
5.3 Set CO2........................................................................................................................... 28
5.4 Set Temperature Offset Compensation.......................................................................... 29
5.5 Set CO2Offset Compensation......................................................................................... 30
5.6 Create a New Experiment .............................................................................................. 31
5.7 Insert a CEROtube in the CERO....................................................................................... 32
5.8 Start an Existing Experiment .......................................................................................... 33
5.9 Follow-up a Running Experiments.................................................................................. 34
5.10 View the Output of a Previous Experiment .................................................................. 34
5.11 Export the Output Files of a Previous Experiment ........................................................ 35
5.12 Delete the Output Files of Previous Experiments ......................................................... 35
5.13 Modify System Settings................................................................................................ 36
5.13.1 CO2Alarm ....................................................................................................................36
5.13.2 Tolerance of the Out of Range Notification (CO2and Temperature)..........................36
5.13.3 Cover Drive Notification..............................................................................................37
5.13.4 Date and Time .............................................................................................................37
5.13.5 Message Level .............................................................................................................37
5.13.6 Graph Scaling ..............................................................................................................37
5.13.7 Brightness of the Touch Screen (TFT Brightness)........................................................37
5.13.8 Network Settings.........................................................................................................38
5.14 Update Software.......................................................................................................... 39

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5.15 Delete all Outputs (Format Data Partition) ...................................................................39
5.16 Format USB Stick ..........................................................................................................39
5.17 Transfer Experiments between two CEROs...................................................................39
6. PH-READER.....................................................................................................40
6.1 General Information pH-Reader .....................................................................................40
6.1.1 Intended Use ................................................................................................................ 40
6.1.2 Precautions when installing the Reader ....................................................................... 40
6.2 Description pH-Reader ...................................................................................................40
6.2.1 The Reader ................................................................................................................... 40
6.2.2 Principle of the pH Monitoring..................................................................................... 41
6.2.3 The pH Monitoring Step by Step .................................................................................. 42
6.2.4 Example Outputs .......................................................................................................... 42
6.3 Installation Procedure pH-Reader...................................................................................44
6.4 Operation with pH-Reader .............................................................................................46
6.4.1 Enable Reader............................................................................................................... 46
6.4.2 Set General Reader Settings ......................................................................................... 46
6.4.3 Enable the pH or the Color Mode................................................................................. 47
6.4.4 Adjust the Settings for the Color Monitoring ............................................................... 48
6.4.5 Adjust the Settings for the pH Monitoring ................................................................... 48
6.4.6 Start an Existing Experiment with enabled pH-Reader ................................................ 49
6.4.7 Follow-up the pH Monitoring of a Running Experiment .............................................. 52
6.4.8 Single Point Measurement during a Running Experiment ........................................... 52
7. MAINTENANCE ...............................................................................................53
7.1 Weekly Maintenance......................................................................................................53
7.2 Decontamination of the CERO ........................................................................................53
7.3 Remove and Re-Install the Front Cover ..........................................................................54
7.4 Remove and Re-Install the Convection Channel .............................................................55
7.5 Replace Hepa Filters .......................................................................................................55
7.6 Replace Tube Holders .....................................................................................................56
7.7 Maintenance pH-Reader.................................................................................................56
7.7.1 Weekly Maintenance pH-Reader ................................................................................. 56
7.7.2 Decontamination pH-Reader........................................................................................ 57
7.8 Check the Memory Space Available................................................................................57
7.9 Yearly Maintenance........................................................................................................57
8. TROUBLESHOOTING .......................................................................................58
8.1 Temperature ..................................................................................................................58
8.1.1 Temperature Too High ................................................................................................. 58
8.1.2 Temperature Too Low .................................................................................................. 58

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8.1.3 Temperature Unstable ..................................................................................................58
8.2 CO2Level........................................................................................................................ 59
8.2.1 CO2Alarm Triggered......................................................................................................59
8.2.2 CO2Level Too High ........................................................................................................59
8.2.3 CO2Level Too Low.........................................................................................................60
8.2.4 CO2Level Unstable ........................................................................................................60
8.2.5 CO2Consumption Too High...........................................................................................60
8.3 Tube Rotation Interrupted ............................................................................................. 61
8.4 Convection Channel Interrupted .................................................................................... 61
8.5 Malfunctioning User Interface ....................................................................................... 61
8.6 Sliding Top Cover Does Not Open .................................................................................. 61
8.7 Getting Technical Support.............................................................................................. 62
8.8 System Information ....................................................................................................... 63
8.9 Export System Data........................................................................................................ 63
8.10 Returning an Instrument for Repair ............................................................................. 64
9. TECHNICAL SPECIFICATIONS........................................................................... 65
9.1 Instrument ..................................................................................................................... 65
9.2 pH-Reader...................................................................................................................... 66
10. ORDERING INFORMATION............................................................................ 67
10.1 Instrument ................................................................................................................... 67
10.2 Consumables................................................................................................................ 67
11. GLOSSARY .................................................................................................... 68

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PROLOGUE
I. Revision History
Version
Revision Date
2018-12
December 2018
© Copyright 2018, OLS, OMNI Life Science GmbH & Co. KG. All rights reserved.
Information in this document is subject to change without notice. No part of this document
may be reproduced or transmitted in any form or by any means, electronic or mechanical, for
any purpose, without the express written permission of OLS OMNI Life Science.
Questions or comments regarding the contents of this Operators Guide can be directed to the
address below.
OMNI Life Science GmbH & Co. KG
Karl-Ferdinand-Braun-Str. 2
28359 Bremen
Telefone: 0421 | 27 61 69-0
e-mail: [email protected]
Online Support
CERO online is available on www.ols-bio.de.
+direct online contact
+manuals, literature & applications
+video manuals
+tips & tricks
+troubleshooting
Every effort has been made to ensure that all the information contained in the CERO Operators
Guide is correct at the time of printing. However, OLS OMNI Life Science GmbH & Co. KG
reserves the right to make any changes necessary without notice as part of ongoing product
development.
.
II. Contact Addresses
Manufacturer OMNI Life Science GmbH & Co. KG
Karl-Ferdinand-Braun-Str. 2
28359 Bremen
Fone: +49-421 | 27 61 69-0
e-mail: [e-mail]
www.ols-bio.de

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III. Declaration of Conformity
The instrument meets the requirements laid down in Council Directive
2014/30/EC relating to “Electromagnetic Compatibility”and Council
Directive 2014/35/EC relating to “Low Voltage Equipment”. The following
standards were applied: EN 61010-1:2001 and EN 61010-2-081:2002
(Safety); EN 61326-:2013 (EMC Emision, Class B) and EN61326-1:2013
(EMC Immunity)
IV. Warranty
The warranty conditions are specified in the sales contract. Contact your OLS representative for
further information. Any unauthorized modification of the instrument invalidates the
guarantee and service contract.

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1. GENERAL INFORMATION
1.1 Intended Use
The CERO is a bench-top incubator and bioreactor hybrid capable of handling four independent
high-density micro carrier or suspension cell cultures.
At the present time the CERO is classified as a general laboratory instrument and is not an in
vitro diagnostic device: for research only.
1.2 About this Operators Guide
This Operators Guide will help you to operate the CERO correctly and safely. To achieve that
goal, this Operators Guide will describe the different components of the CERO and how they
function. The Operators Guide describes both the hardware and software of the CERO in a
depth enabling you to operate the instrument.
After introducing you to the various parts of the CERO, we will show you step by step how to
operate the instrument. When you have read this Operators Guide, you should be capable of
operating the CERO.
Warnings are included in this Operators Guide to emphasize important and
critical instructions. They are printed in bold italics, and begin with the word
“ATTENTION” accompanied by the symbol on the left.
Biohazard risk warnings are printed in bold italics, and begin with the word
“BIOHAZARD” accompanied by the symbol on the left.
Important Notes are included in this Operators Guide to emphasize important
instructions. They are printed in italics, and begin with the word “IMPORTANT
NOTE” accompanied by the symbol on the left.
Notes and additional information, that can be helpful for your work with the
CERO are printed in italics, and begin with the word “NOTE” accompanied by
the symbol on the left.
1.3 Operation
The operator of the CERO must be familiar with the instructions lined out in this Operators
Guide. The procedures contained within this Operators Guide have been tested by the
manufacturer and are deemed to be fully functional. Any deviation from the procedures given
here could lead to erroneous results or malfunction, and may be dangerous.

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2. SAFETY PRECAUTIONS AND HAZARDS
The following section describes the main safety considerations in operating this product and
the main hazards involved.
ATTENTION
Read the following safety notices carefully before installing and operating
the CERO.
2.1 General Precautions
2.1.1 Instrument
For safe and correct use of the CERO, it is essential that both operating and service personnel
follow generally accepted safety procedures as well as the safety instructions in this document,
the CERO Operators Guide.
The instrument should be positioned in the laboratory in a way permitting personnel to access
the front and sides of the instrument for operation and maintenance. Accordingly, to calculate
how much space is needed, consider the dimensions of the instrument, the length of the power
cable, the distance to the CO2 supply and the space for a person to move and work comfortably
(please refer to the Chapter 9 Technical Specifications).
Cleaning, dismantling, and maintenance should only be performed by personnel trained in such
work, and who are aware of the possible dangers involved. Only certified technicians are
authorized to perform mechanical maintenance on the CERO.
Necessary precautions should be taken when transporting the instrument. For repair or
shipment, all mechanical parts must be put in their rest positions. The CERO must be sent back
in the original reusable packaging box. There must not be any labware in the CERO during
transportation. Any CERO sent away for repair must also be decontaminated (see Chapter 8.9).
Only original CERO-approved parts and accessories may be used with the CERO. Any alterations
or modifications to your system may be dangerous and will void your warranty.
2.1.2 Operating the CERO
When using the CERO, Good Laboratory Practices (GLP) must be observed. Wear suitable
protective clothing, safety glasses and protective gloves, particularly when dealing with a
malfunction of the instrument where the risk of contamination exists.
During CERO operation, stand clear of any moving parts. Do not attempt to block the sliding
cover while opening or closing it.
In case of incidental spillage, the instrument must be thoroughly decontaminated by spraying
it or wiping it down with the appropriate disinfectant taking into account the nature of the spill
and the necessary safety precautions.
Do not attempt to move the unit while in operation.

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2.2 Hazards of CO2
ATTENTION
CO2is a colourless, odourless and tasteless gas. Because CO2displaces
oxygen it represents a health risk as asphyxiant.
The user is responsible for providing safe and professional CO2supply to the CERO. Do not
connect the CERO to a malfunctioning CO2supply and regularly check the integrity of your CO2
supply. Make sure that the CO2supply is properly fixed to the CERO.
Always wear a personal detection and warning device while working in the room where the
CERO has been installed.
Always turn off the CO2supply when the CERO is not running.
Do not attempt to move the unit while connected to the CO2supply.
2.3 Electrical Safety Precautions
The CERO must first be switched off and disconnected from the main electrical supply before
removing any mechanical or electrical component.
Do not connect the unit to a power source of any other voltage or frequency beyond the range
stated on the power rating in the Technical Specifications (see Chapter 9).
Avoid damaging the power cord. Do not bend it excessively, step on it, place heavy objects on
it. A damaged cord can easily become a shock or fire hazard. Never use a power cord after it
has become damaged.
Only connect the CERO to an earth-grounded outlet.
2.4 Biohazard Precautions
BIOHAZARD
The CERO does not provide any user protection against biohazards, such as
the one provided by a class II cabinet.
Selection of the appropriate biological safety level and implementation of the required
biosafety measures for working with the CERO is the sole responsibility of the user.
If the CERO becomes contaminated with biohazardous, radioactive or chemical material, it
should be cleaned (see also Chapter 7). Failure to observe and carry out the maintenance
procedures given may impair the reliability and correct functioning of the CERO.
If working with biohazardous samples observe and carry out the maintenance procedures,
paying particular attention to cleaning and decontamination. Wear gloves during the cleaning
process. Any surfaces on which liquid is spilled must be decontaminated.

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3. PRODUCT DESCRIPTION
This chapter provides a brief overview of the CERO system. See Chapter 4 for complete
installation instructions and Chapter 5 for complete usage instructions.
3.1 Culturing Cells in the CERO
The CERO is a benchtop incubator and bioreactor hybrid supporting aggregate and the cell
culture on the Microcarrier substrates. The primary functions of the CERO are:
- maintenance of the culture environment (e.g. CO2, temperature)
- control of the cell suspension via rotation of the CEROtube
- monitoring the pH of the cell culture medium
Cell culture in the CERO generally consists of three phases:
Inoculation, Culture and Harvesting (figure 1).
Figure 1: Cu lt ur ing Cells in the C ERO –using th e example o f a microc ar ie r culture
The initial inoculation phase supports the aggregation of the cells or the loading of cells on the
microcarrier substrate. The culture phase refers to the expansion of the culture. The harvesting
phase (optional) corresponds to the dissolution of the cells.
3.1.1 Parameters, Protocol and Experiment
Parameters are low level commands which value can be set, in a defined range, by the user via
the user interface.
A protocol consists of a set of commands together with controllable parameters and defines all
necessary actions. There exist 3 types of protocols: inoculation, culture and harvesting, which
are described below.
An experiment consists of the succession of protocols, usually an Inoculation Protocol followed
by a Culture Protocol and a Harvesting Protocol. 4 different experiments, one for each tube,
can be run independently.

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3.1.2 Inoculations Protocol
An Inoculation Protocol defines all necessary actions enabling cell aggregation or adherence/
loading on Microcarriers in the CERO and contains a defined set of parameters controlling the
CEROtube movements (figure 2). The basic CEROtube movement supporting the
homogenisation of the cell suspension is a bidirectional rotation controlled by four parameters:
- Rotation Period: period of rotation into one direction
- Rotation Pause: pause between 2 rotation periods
- Rotation Speed
During inoculation, it is critical to ensure adequate aggregation of cells or adhesion of the cells
to the Microcarriers. Most cell types cannot inoculate under constant agitation conditions.
Thus, two more parameters are controlled in an Inoculation Protocol (figure 2):
- Agitation Period: period during which the CEROtube is undergoing alternating rotation
movements
- Agitation Pause: temporary interruption of the agitation to enhance inoculation
efficiency of certain cell types
Finally, the length of a protocol can be determined with the parameter Protocol Duration.
Figure 2 : S tandard I n o culatio n Protoc ol

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3.1.3 Culture Protocol
After a successful inoculation cell expansion is easily accomplished. The Standard Culture
Protocols (figure 3) consist of the same parameters as the Inoculation Protocol. Since Culture
Protocols usually do not need conditions as gentle as Inoculation Protocols, the default
parameters have different settings e.g. the Agitation Pause is switched off (none). Nevertheless
more gentle conditions can be set with the same functionality as in Inoculation Protocol mode.
Figure 3 : S tandard C ul tu re Pr o tocol
3.1.4 Harvesting Protocol
A standard Harvesting Protocol (figure 4) consists of the following parameters:
- Rotation Period: period of rotation into one direction
- Rotation Pause: pause between 2 rotation periods
- Rotation Speed
For Harvesting no agitation is needed so the parameters Agitation Period and Agitation Pause
were excluded from this protocol.
Figure 4 : S tandard H a rv es ting Protocol

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3.2 Allgemeine Beschreibung des CERO
The CERO consists of 2 basic units (figure 5):
-the Culture Chamber
-the User Interface
Figure 5 : CERO Sys te m Ove rv ie w
The CERO components and accessories are further described in the following sections.
3.3 Culture Chamber
3.3.1 Sealed Culture Chamber
The sealed compact Culture Chamber (figure 6) maintains consistent CO2and temperature
levels during cell expansion. The removable Convection Channel on the left hand side of the
CERO provides proper air circulation and a homogenous environment.
Figure 6: Sealed Culture Chamber
Click-Wheel
Touchscreen
Convection Channel
Sliding Top Cover
Removable Front cover
Open Cover Button
Culture Chamber
User Interface

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The Open Cover Button (figure 6) activates the Top Cover to open and close. This allows
insertion and removal of culture tubes from the Culture Chamber. The Culture Chamber has
also been designed to be very easy to clean. Complete access to the Culture Chamber is made
possible with the easily removable front cover (see Chapter 5 for more information).
3.3.2 CO2and Temperature Control
Integrated with compact controllers the CERO ensures proper control of environmental
temperature and CO2levels (figure 7). The Chamber Access Port on the Top Cover enables
monitoring of the temperature and the CO2level directly in the chamber using traditional
laboratory temperature and CO2analysers.
Figure 7: Co nt ro l of CO2- Le vel
3.3.3 Control of the Cell Suspension
The CERO supports up to four independent and high density cell cultures. The CEROtubes have
been designed to gently homogenise the cell suspension in the CERO through bidirectional
rotation (figure 8).
Figure 8: CE RO tu bes

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3.4 User Interface
The Touch-Screen Interface with Click-Wheel (figure 9) allows for quick navigation and provides
users with intuitive and easy to program cell culture workflows. In addition, culture reports
(including temperature and CO2monitoring) can be viewed real-time, saved and exported for
records of compliance.
Figure 9 : C ontrol Unit

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3.5 CERO Software
The CERO Software is structured in 4 Menus all accessible from the Main Menu (see figure 10):
-the System Menu
-the Experiments Menu
-the Outputs Menu
-the Run Menu
Figure 1 0 : Menu Structu re