Thermo Scientific invitrogen EVOS OSI-2 User manual

For Research Use Only. Not for use in diagnostic procedures.
EVOS™Onstage Incubator (OSI-2)
USER GUIDE
for use with:
EVOS™M5000 Imaging System
EVOS™M7000 Imaging System
Catalog NumberAMC2000
Publication Number MAN0027987
Revision A.0

Life Technologies Corporation | 22025 20th Ave SE Ste. 100 | Bothell, Washington 98021 USA
Revision history:MAN0027987 A.0 (English)
Revision Date Description
A.0 6 March 2023 Initial release.
The information in this guide is subject to change without notice.
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©2023 Thermo Fisher Scientific Inc. All rights reserved.

Contents
■CHAPTER1Productinformation .................................................. 5
Productdescription ............................................................. 5
EVOS™Onstage Incubator(OSI-2) ............................................ 5
Standard itemsincluded ..................................................... 5
Required materials not provided .............................................. 6
Incubatorcomponents ........................................................... 7
Control unit and Environmental chamber ...................................... 7
Control unit (backview) ...................................................... 8
Status LEDmodes .......................................................... 9
Enviromentalchamber ..................................................... 10
■CHAPTER2Methods ............................................................. 12
Set up the EVOS™Onstage Incubator(OSI-2) ...................................... 12
Install the EVOS™Onstage Incubator (OSI-2)  ................................. 12
Set up the EVOS™Onstage Incubator (OSI-2) for operation ..................... 15
Use the Onstage Incubator with the EVOS™M5000 Imaging System ................. 19
Power on the EVOS™Onstage Incubator(OSI-2) ............................... 19
Optional: Set Ambient Temperature Oset .................................... 22
Configure gasinputs ....................................................... 23
Calibrate incubator oxygensensor ........................................... 24
Use the Onstage Incubator with the EVOS™M7000 Imaging System ................. 27
Power on the EVOS™Onstage Incubator(OSI-2) ............................... 27
Configure gasinputs ....................................................... 29
Calibrate oxygensensor .................................................... 31
■CHAPTER3Troubleshooting ..................................................... 33
Troubleshooting ................................................................ 33
■APPENDIXASpecifications ...................................................... 35
Physical characteristics ......................................................... 35
Hardware ..................................................................... 35
EVOS™Onstage Incubator (OSI-2) User Guide 3

■APPENDIXBCompatible vessel holders and stage plates ................... 36
EVOS™Onstage Incubator (OSI-2)-compatible vessel holders and stage plates ....... 36
■APPENDIXCSafety ............................................................... 39
Symbols on thisinstrument ..................................................... 39
Standard safetysymbols ................................................... 39
Location of safetylabels .................................................... 40
Control and connectionsymbols ............................................. 41
Conformitysymbols ........................................................ 41
Safety information for instruments not manufactured by Thermo FisherScientific ...... 42
Instrumentsafety .............................................................. 42
General ................................................................... 42
Electricalsafety ........................................................... 42
Safety requirements for EVOS™Onstage Incubator(OSI-2) ...................... 43
Cleaning anddecontamination .............................................. 44
Instrument component and accessorydisposal ................................ 44
Safety and electromagnetic compatibility (EMC) standards .......................... 45
Safety standards .......................................................... 45
EMC standards ............................................................ 45
Environmental design standards ............................................. 46
Chemicalsafety ................................................................ 46
Chemical wastesafety .......................................................... 47
Chemical waste hazard ..................................................... 47
Chemical waste safetyguidelines ............................................ 47
Wastedisposal ............................................................ 47
Biological hazardsafety ......................................................... 48
■APPENDIXDDocumentation and support ...................................... 49
Customer and technical support ................................................. 49
Limited product warranty ........................................................ 49
Contents
4EVOS™Onstage Incubator (OSI-2) User Guide

Product information
IMPORTANT! Before using this product, read and understand the information in the “Safety” appendix
in this document.
Product description
EVOS™Onstage Incubator (OSI-2)
The EVOS™Onstage Incubator (OSI-2) is an optional accessory for the EVOS™M5000 and EVOS™
M7000 Imaging Systems that enables the incubation of cells at user-defined temperature, humidity, and
gas concentration (O2, CO2), allowing the capture of images from the same sample over long periods of
time and recording of time lapse movies.
The EVOS™Onstage Incubator (OSI-2) consists of a stagetop environmental chamber that is placed on
the X‑Y stage of the imaging system and a separate control unit that supplies the power and humidified
gas (air or air-CO2premix, CO2-only, and N2-only). The second generation EVOS™Onstage Incubator
contains an on‑board air compressor that enables the use of on‑board air instead of a separate air tank.
The Onstage Incubator (OSI-2) is controlled by the same software and user interface that controls the
imaging system.
Standard items included
•Stagetop Environmental Chamber
•Control Unit
•Master Stage Plate (also available separately as EVOS™Onstage Master Plate,
Cat.No.AMEPVH106)
•Vessel holder for multiwell plates (also available separately as EVOS™Onstage Vessel Holder,
Multiwell Plates, Cat.No.AMEPVH028)
•Light shield and light shield cover
•Sensor data cable with 6-pin connector
•Cable, USB2.0A‑to‑B, 180cm (6ft)
•Heated hose with temperature control, 180 cm (6 ft) (also available separately as EVOS™Onstage
Incubator Hose, Cat.No.AMEP4728)
•Gas line, 1/8 in. ID, ¼ in. OD (also available separately as EVOS™Onstage Incubator Gas Line,
Cat.No.AMEP4732, for an extra 25ft)
•Push-to-connect gas line adaptor, ¼in. NPT male (3each)
•Standard-head open-end wrench
•Hex screw driver (2mm)
1
EVOS™Onstage Incubator (OSI-2) User Guide 5

•Power Cord, Type A (North America)
Note: A country-specific power cord must be ordered separately in regions not using the TypeA
power plug.
Required materials not provided
•CO2gas or CO2/air premix for CO2regulation
•N2gas for O2regulation (if performing hypoxia experiments)
•Multistage regulators capable of providing continuous 40–50 psi output pressure for all gas sources
used with the OSI-2. The regulators must have a ¼ in. NPT female output; if the regulator has a
¼ in. NPT male output, a male-to-female adaptor is needed.
Chapter1Product information
Product description
1
6EVOS™Onstage Incubator (OSI-2) User Guide

Incubator components
Control unit and Environmental chamber
2
4
7
1
6
3
5
Figure1EVOS™Onstage Incubator (OSI-2) Control unit and Environmental chamber
1Control unit
2Heated hose
3Hose heater connector
4Water reservoir door
5Water reservoir door release
6Environmental chamber
7Sensor data cable with 6‑pin connector
Chapter1Product information
Incubator components 1
EVOS™Onstage Incubator (OSI-2) User Guide 7

Control unit (back view)
Figure2Back view of the OSI-2 Control unit showing the locations of the gas ports, input jacks, and the
power switch
1Port 1: Air or air‑CO2premix
2Port 2: CO2
3Port 3: N2
4Power switch
5Power input jack
66‑pin sensor data cable jack
7USB control cable jack
Chapter1Product information
Incubator components
1
8EVOS™Onstage Incubator (OSI-2) User Guide

Status LED modes
The status indicator LED on the OSI-2 control unit identifies the status of the incubator.
Common Callouts and Arrows
1. Copy-paste a callout or arrow to use in this SVG.
Note: If you need more advanced callouts or arrows
use the TechComm_Inkscape_Callout&Arrow_Libary.
3. Delete this text, this rectangle, and unused
callouts, arrows, or other SVG elements
before adding this SVG to the repository.
2. Edit number and/or line-length, as needed.
1 1
1
1
1
Figure3OSI-2 status indicator LED on the control unit
1Status indicator LED
LED color LED mode Incubator status
Blue Solid The OSI-2 is powered on and connected to the computer, but no
functions have been enabled.
Blue Blinking The temperature/humidity/gas concentration features have been
enabled and the system is warming up.
Blue Slow blink–
fadein/fadeout
All temperature/humidity/gas concentration targets are met and being
maintained.
Amber Blinking The OSI-2 has deviated from the temperature/humidity/gas
concentration target.
Red Blinking The USB cable is connected, but the power switch is o or no power
supply is connected.
Chapter1Product information
Incubator components 1
EVOS™Onstage Incubator (OSI-2) User Guide 9

Enviromental chamber
The environmental chamber of the EVOS™Onstage Incubator (OSI-2) consists of the incubator
chamber, the vessel holder/adaptor, the heated glass lid, the light shield, and the light shield cover.
The environmental chamber sits on the onstage incubator master stage plate that is attached to the X-Y
stage of the imaging system.
6
5
3
2
1
4
7
Figure4Components of the Environmental chamber
1Light shield cover
2Light shield
3Heated glass lid
4Vessel holder/adaptor
5Incubator chamber
6Master stage plate
7Perfusion ports
Chapter1Product information
Incubator components
1
10 EVOS™Onstage Incubator (OSI-2) User Guide

Common Callouts and Arrows
1. Copy-paste a callout or arrow to use in this SVG.
Note: If you need more advanced callouts or arrows
use the TechComm_Inkscape_Callout&Arrow_Libary.
3. Delete this text, this rectangle, and unused
callouts, arrows, or other SVG elements
before adding this SVG to the repository.
2. Edit number and/or line-length, as needed.
1 1
1
1
1
1
Figure5Bottom view of the Environmental chamber lid showing the lid contacts
1Environmental chamber lid contacts
Chapter1Product information
Incubator components 1
EVOS™Onstage Incubator (OSI-2) User Guide 11

Methods
Set up the EVOS™Onstage Incubator (OSI-2)
IMPORTANT! Before using this product, read and understand the information in the “Safety” appendix
in this document.
Install the EVOS™Onstage Incubator (OSI-2)
Assemble the Environmental chamber
1. Place the incubator chamber on the onstage incubator master plate, then secure it in place with
the four 2.0‑mm hex screws.
1
2
3
1Incubator chamber
2Onstage incubator master plate
32.0‑mm hex screws
2. Attach the vessel holder/adaptor to the incubator chamber using the four thumb screws.
1
3
2
4
1Vessel holder/adaptor
2Incubator chamber
3Onstage incubator master plate
4Thumb screws
2
12 EVOS™Onstage Incubator (OSI-2) User Guide

Install the Onstage Incubator Master Plate
Note: The images in the following steps show the installation of the master plate on the EVOS™M5000
Imaging System X‑Y stage. However, the steps to install the master plate on the EVOS™M7000 Imaging
System are identical to the steps for the EVOS™M5000 Imaging System.
1. Unscrew the four 3.0‑mm screws on the base plate, then remove the base plate from the
X‑Y stage. If necessary, unscrew and remove the vessel holder/adaptor before removing the base
plate.
3
1
2
1Base plate
2X‑Y stage
33.0‑mm screw
Chapter2Methods
Set up the EVOS™Onstage Incubator (OSI-2) 2
EVOS™Onstage Incubator (OSI-2) User Guide 13

2. Secure the onstage incubator master plate with the environmental chamber to the X‑Ystage with
the four thumb screws.
2
1
3
4
5
1Environmental chamber
2Onstage incubator master plate
3X‑Y stage
4Perfusion ports
5Thumb screw
IMPORTANT! If you plan to use the perfusion ports on the environmental chamber, install the
master plate such that the perfusion ports face towards the front of the system (as shown in the
image above). Otherwise, install the master plate such that the perfusion ports face the back of the
system.
Chapter2Methods
Set up the EVOS™Onstage Incubator (OSI-2)
2
14 EVOS™Onstage Incubator (OSI-2) User Guide

Set up the EVOS™Onstage Incubator (OSI-2) for operation
Follow the procedures below to set up the EVOS™Onstage Incubator (OSI-2) for operation. For the
locations of various input jacks and gas ports, refer to “Control unit (back view)” on page8.
IMPORTANT! Do not position the control unit so that it is dicult to turn o the main power switch
(see page 8 for the location of the power switch). In case of an instrument malfunction, turn the main
power switch to the OFF position and disconnect the power cord from the outlet.
Connect the gas lines
1. Connect each gas line to the appropriate gas tank via the PTC (push-to-connect) connectors
threaded into the regulator. To do this, push the tubing into the PTC connector until it clicks into
place. Pull on the tubing slightly to ensure a tight connection; the tubing should not come out.
2. Attach the gas lines to the control unit via the PTC connectors for the appropriate gas intake port.
•If using external air or premixed air, attach to Port 1: Air.
•If using CO2, attach to Port 2: CO2.
•For nitrogen or oxygen displacement, attach to Port 3: N2.
Note: A combination of air and CO2gases is needed to perform normoxic live cell imaging with
the OSI-2, which can be accomplished with one of these configurations:
·OSI-2 on-board air compressor + CO2tank
·Premix gas tank containing air + CO2
·Compressed air tank + CO2tank
The use of a pure O2tank is not recommended.
To perform hypoxic cell imaging, the following tank configuration is recommended:
·N2tank + CO2tank
Chapter2Methods
Set up the EVOS™Onstage Incubator (OSI-2) 2
EVOS™Onstage Incubator (OSI-2) User Guide 15

Connect the power and system control lines
1. Plug the power cord into the power input jack on the control unit and the outlet.
2. Plug the USB cable into the USB control cable jack on the control unit and the USB port on the
imaging system computer.
3. Plug the 6-pin sensor data cable to the stagetop environmental chamber and the appropriate input
jack on the control unit.
Set up the control unit
1. Assemble the water reservoir and add warm water (approximately 50°C) to the max fill line through
the fill hole. Distilled water is recommended to avoid contaminants.
IMPORTANT! Do not overfill the water reservoir.
1
1Reservoir fill holes
2. Place the water reservoir into the control unit with the fill holes to the front and close the lid.
1
2
3
1EVOS™Onstage Incubator (OSI-2) control unit
2Water reservoir
3Water reservoir door release
Chapter2Methods
Set up the EVOS™Onstage Incubator (OSI-2)
2
16 EVOS™Onstage Incubator (OSI-2) User Guide

Set up the Environmental chamber for operation
1. Place the heated glass lid with the no-fog glass window on the incubator chamber. The heated
glass lid is guided and attached in its place by the two magnets on its rim.
1
2
3
1Heated glass lid
2Incubator chamber
3Onstage incubator master
plate
2. Place the light shield with tinted plastic window on top of the heated glass lid.
Note: Use of the light shield is strongly recommended for fluorescence imaging applications.
1
2
3
4
1Light shield cover
2Light shield
3Heated glass lid
4Incubator chamber
Chapter2Methods
Set up the EVOS™Onstage Incubator (OSI-2) 2
EVOS™Onstage Incubator (OSI-2) User Guide 17

3. If desired, place the light shield cover on the light shield for fluorescence imaging applications.
Note: The light shield cover completely blocks any ambient light from entering the environmental
chamber and improves image quality in fluorescence imaging applications.
4. Attach the heated hose between the stagetop environmental chamber and the control unit.
5. Plug the hose heater cable from the control unit into the connector on the heated hose.
Note: The following image shows the environmental chamber set up for operation on the EVOS™
M5000 Imaging System. However, the steps to set up the environmental chamber on the EVOS™
M7000 Imaging System are identical to the steps for setup on the EVOS™M5000 Imaging System.
Common Callouts and Arrows
1. Copy-paste a callout or arrow to use in this SVG.
Note: If you need more advanced callouts or arrows
use the TechComm_Inkscape_Callout&Arrow_Libary.
3. Delete this text, this rectangle, and unused
callouts, arrows, or other SVG elements
before adding this SVG to the repository.
2. Edit number and/or line-length, as needed.
1 1
1
1
1
2
34 5
7
6
1Imaging system
2X‑Y stage
3Stagetop environmental chamber
4Sensor data cable
5Heated hose
6Hose heater cable
7Control unit
Chapter2Methods
Set up the EVOS™Onstage Incubator (OSI-2)
2
18 EVOS™Onstage Incubator (OSI-2) User Guide

Use the Onstage Incubator with the EVOS™M5000 Imaging
System
Note: For EVOS™M7000 Imaging System instructions, see “Use the Onstage Incubator with the
EVOS™M7000 Imaging System” on page27.
IMPORTANT! Before using the EVOS™Onstage Incubator (OSI-2) in experiments, ensure that:
·The gas inputs have been configured (see “Configure gas inputs” on page23).
·The oxygen sensor has been calibrated (see “Calibrate incubator oxygen sensor” on page24).
Power on the EVOS™Onstage Incubator (OSI-2)
1. Power on the EVOS™M5000 Imaging System as described in the EVOS™M5000 Imaging System
User Guide (Pub. No. MAN0017563).
2. Power on the EVOS™Onstage Incubator (OSI-2) control unit.
3. Start the EVOS™M5000 Software. The software recognizes the Onstage Incubator on connection.
4. On the Capture tab, click Automate, then select Incubator to open the Incubator Control
window.

Common Callouts and Arrows
1. Copy-paste a callout or arrow to use in this SVG.
Note: If you need more advanced callouts or arrows
use the TechComm_Inkscape_Callout&Arrow_Libary.
3. Delete this text, this rectangle, and unused
callouts, arrows, or other SVG elements
before adding this SVG to the repository.
2. Edit number and/or line-length, as needed.
1 1
1
1

Chapter2Methods
Use the Onstage Incubator with the EVOS™M5000 Imaging System 2
EVOS™Onstage Incubator (OSI-2) User Guide 19

Note: Alternatively, go to the Settings tab, click Incubator to expand the Incubator panel, then
click Incubator Control to open the Incubator Control window.

Common Callouts and Arrows
1. Copy-paste a callout or arrow to use in this SVG.
Note: If you need more advanced callouts or arrows
use the TechComm_Inkscape_Callout&Arrow_Libary.
3. Delete this text, this rectangle, and unused
callouts, arrows, or other SVG elements
before adding this SVG to the repository.
2. Edit number and/or line-length, as needed.
1 1
1
1

5. Select Enable to initiate the control of the incubator.
6. If you are using the incubator for the first time or if the configuration of the gas inputs has changed,
configure the gas inputs (see “Configure gas inputs” on page23).
Chapter2Methods
Use the Onstage Incubator with the EVOS™M5000 Imaging System
2
20 EVOS™Onstage Incubator (OSI-2) User Guide