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ibidi Gas Incubation System – Silver Line

ibidi

Gas Mixer

Gas Mixer

Humidification

Stage Top Incubator

Humidity

sensor

Valve

regulation

Gas mixture

(humidified)

Dry gas

Gas mixture

(dry)

Super-humidified gas

“100%+” RH

Feedback control

Air N

2

O

2

Dry gas mix
Humidified gas mix

Humidified and warm

gas mixture

ibidi

Temperature

Controller

Figure 2: Schematic view of the ibidi Stage Top Incubation System, combining the ibidi Heating
System with the ibidi Gas Incubation System.

In combination with the ibidi Heating System, it supplies complete incubator conditions for your live
cell imaging directly on the microscope stage.

Humidity in ambient air can lead to condensation on all surfaces, especially on the lids of cell culture
vessels. If these surfaces are within the optical pathway, small water droplets will cause light scat-
tering. This diminishes the optical quality of transmitted light microscopy (i.e., phase contrast). To
ensure the highest image quality, condensation on any surface should be prevented during live cell
imaging experiments.

The independently controlled Heated Lid of the ibidi Heating System prevents from condensation
during live cell imaging. By heating the lid to a temperature higher than the plate, a vertical tem-
perature gradient is created. This gradient, and active humidity control prevent the formation of
condensation on the lid of the cell culture vessel.

34 °C

Unheated Lid

Heated Plate

37

 

°C

ibidi solution:

 

Heated Lid

40 °C

Heated Plate

37

 

°C

Figure 3: The ibidi Heating System prevents condensation on the cell culture vessel lid, as well as on
the Heated Lid itself.

Version 1.0 (June 1, 2022)

16

Instruction Manual

Summary of Contents for 12110

Page 1: ...lide Dish Silver Line ibidi Gas Mixer CO 2 O2 Flow I 9 9 S 10 0 Run CO2 Hum 5 1 5 0 75 2 80 0 Setup ibidi Gas Mixer CO2 O2 Flow I 9 9 S 10 0 Run CO2 Hum 5 1 5 0 75 2 80 0 Setup 12110 12720 12722 12111 11921 ibidi Gas Incubation System CO2 Silver Line 11923 ibidi Gas Incubation System CO2 O2 Silver Line com ...

Page 2: ...ibidi Gas Incubation System Silver Line Version 1 0 June 1 2022 2 Instruction Manual ...

Page 3: ...ion System Silver Line Contact ibidi GmbH Lochhamer Schlag 11 82166 Gra felfing Germany Phone 49 89 520 46 17 0 Fax 49 89 520 46 17 59 E mail info ibidi de Internet ibidi com Instruction Manual 3 Version 1 0 June 1 2022 ...

Page 4: ...ne 12 1 9 Repairing the ibidi Gas Incubation System Silver Line 13 1 10 Waste Disposal WEEE RoHS Compliance Statement 13 1 10 1 EU Directive WEEE 13 1 10 2 EU Directive RoHS 13 1 11 Regulatory Statement 14 2 Intended Use 15 3 Principle 15 4 Equipment 17 4 1 Components of the ibidi Gas Incubation System Silver Line 17 4 2 Gas Mixer CO2 or CO2 O2 18 4 3 Humidifying Column 20 4 4 Heated Tubing Packag...

Page 5: ... 31 5 4 1 Run Stop Mode 31 5 4 2 Incubation Parameters 32 5 4 3 Setup Menu 33 6 IncuControl Software 35 7 Maintenance 36 7 1 Disinfection and Cleaning 36 7 1 1 Wiping the Outer Surface 36 7 1 2 Cleaning the Column s Inner Face 36 7 2 Influence of Surrounding Temperature and Airflow 36 8 Troubleshooting 37 8 1 Controller Display Errors 37 8 2 Flow Readings 37 8 3 CO2 Channel Readings 38 8 4 O2 Chan...

Page 6: ...Silver Line with the ibidi Heating System 4 Slides Silver Line 12130 12131 or ibidi Heating System Multiwell Plate Silver Line 12150 12724 12726 12151 please refer to the instruction manual for use with the ibidi Heating System 4 Slides Silver Line and ibidi Heating System Multiwell Plate Silver Line 1 2 Safety Symbols Note that the signal words WARNING CAUTION and NOTE have specific meanings in t...

Page 7: ... 4 Specifications Table 1 Specifications of the ibidi Gas Incubation System Silver Line Electrical Specifications Power Supply Protection class I International protection marking IEC 60529 IP 20 Overvoltage category II External power supply AC 100 240 V 50 60 Hz 1 4 A Input line voltage Gas Mixer DC 24 V 5 A 120 W Output voltage to Humidifying Column DC 24 V max 3 6 A Operating Conditions Operatin...

Page 8: ...tings Gas Mixer input ID 4 mm OD 6 mm Gas Mixer output and Humidify ing Column connection ID 2 5 mm OD 4 mm Connection to the Heated Lid push in fitting ID 2 5 mm OD 4 mm Gas Input Requirements Compressed air Purity class ISO 8573 1 2010 1 2 1 1 bar 14 5 psi optimum 0 8 1 2 bar 11 6 17 4 psi Carbon dioxide CO2 Purity 99 CO2 1 bar 14 5 psi optimum 0 8 1 2 bar 11 6 17 4 psi Nitrogen N2 Purity 99 N2 ...

Page 9: ...ime is three years under continuous operation only for the CO2 O2 version Humidity Control Humidity sensor type Sensirion SHT35 DIS Control range 20 99 lower limit dependent on environment and gas supply min set humidity is 0 Sensor accuracy 2 5 max Resolution 0 1 Humidifying Column Temperature Control Control range 45 C to 55 C Accuracy 2 C Resolution 0 1 only controllable on the Gas Mixer not in...

Page 10: ...adequate specifications By violating these instructions you risk electric shock and fire Only use extension cables that have a protective ground wire Do not operate the ibidi Gas Incubation System Silver Line under conditions that pose a risk of explosion implosion or the release of gases Only operate the ibidi Gas Incubation System Silver Line with ultra pure water Do not place flammable solids l...

Page 11: ...uency The ibidi Gas Incubation System Silver Line might not function properly when located near a strong magnetic field or high frequency source Avoid vibrations from vacuum pumps centrifuges electric motors processing equipment and machine tools Avoid dust and corrosive gas Do not install the ibidi Gas Incubation System Silver Line where it could be exposed to high levels of dust or to outside ai...

Page 12: ...urrounding gases 7 Accidents beyond ibidi s control including natural disasters This warranty does not cover consumables such as cell culture chambers and dishes tubes fluidic connectors reagents etc The warranty for all parts supplied and repairs provided under this warranty expires on the warranty expiration date of the original product 1 8 Transporting the ibidi Gas Incubation System Silver Lin...

Page 13: ...ous Substances RoHS 1 10 1 EU Directive WEEE The ibidi Gas Incubation System Silver Line must be disposed of in compliance with the WEEE Directive 2012 19 EC This symbol on the product is in accordance with the European Union s Waste Electrical and Elec tronic Equipment WEEE Directive The symbol indicates that this product must be recycled dis posed of separately from other household waste It is t...

Page 14: ... declaration Laborgerät laboratory equipment ibidi Gas Mixer ibiGM2 XXX with ibidi accessories Der oben beschriebene Gegenstand erfüllt die Vorschriften der Richtlinie 2011 65 EU vom 08 Juni 2011 zur Beschränkung der Verwendung bestimmter gefährlicher Stoffe in Elektro und Elektronikgeräten The object of the declaration described above is in conformity with Directive 2011 65 EU of 8 June 2011 on t...

Page 15: ...e ibidi Heating System Silver Line 3 Principle Physiological Conditions in Live Cell Imaging Cells react sensitively to changes in their environment Factors such as temperature humidity and CO2 O2 levels significantly influence the outcome of cell culture assays In order to achieve biologi cally relevant and reproducible results it is crucial to maintain optimal conditions on the microscope stage ...

Page 16: ...ture vessels If these surfaces are within the optical pathway small water droplets will cause light scat tering This diminishes the optical quality of transmitted light microscopy i e phase contrast To ensure the highest image quality condensation on any surface should be prevented during live cell imaging experiments The independently controlled Heated Lid of the ibidi Heating System prevents fro...

Page 17: ...ne Cat No Description Components 11921 ibidi Gas Incubation System Silver Line for CO2 Gas Mixer CO2 Humidifying Column Heated Tubing Package Gas Tubing for CO2 and Air USB cable Power cable USB flash drive 11923 ibidi Gas Incubation System Silver Line for CO2 and O2 Gas Mixer CO2 O2 Humidifying Column Heated Tubing Package Gas Tubing for CO2 and Air USB cable Power cable USB flash drive Gas Tubin...

Page 18: ...ket USB flash drive with IncuControl software Only for the CO2 O2 Version 4 2 Gas Mixer CO2 or CO2 O2 The Gas Mixer is the gas mixing and control unit of the Gas Incubation System There are two versions of the Gas Mixer controlling either the amount of CO2 or the amount of both CO2 and O2 Using the CO2 O2 version hypoxia conditions can be created by replacing O2 with nitrogen N2 In addition to the...

Page 19: ... very high please contact ibidi technical support if blinking persists LED Humidifier blinking Possible channel error please contact ibidi tech nical support On the back of the Gas Mixer all connections for cables and tubing are provided Figure 5 In the CO2 version the N2 port is closed inside the instrument and thus non functional The Gas Incubation System takes as input pure CO2 gas and pressuri...

Page 20: ...as connections input and output as well as an electric plug for the heating of the column connection see Section 5 1 2 On top of the column is an opening for re filling it with ultra pure water filling see Section 5 2 The maximum 130 ml and minimum 50 ml levels are indicated by the red labels inside the column see Figure 6 CAUTION The Humidifying Column base heats up to 50 C 122 F when in operatio...

Page 21: ...ubing The wire is split off of the electrical cable under the insulating jacket enclosing the tubing The exact connection of the tubing is shown in Figure 7 A B C D E F G H I 1 2 Humidifying Column Incubation Chamber Gas Mixer J Figure 7 Heated Tubing Package Electrical cable for the heating of the Humidifying Column A insulating jacket B Mixed gas output white and yellow C and D heated wire split...

Page 22: ...ty inside the Heated Lid The humidity sensor is connected with the Heated Plate via the electrical connection pins of the Heated Lid The cable of the Heated Plate guides the information of the humidity sensor to the Temperature Controller For connection with the Gas Mixer an extra bridge cable is needed see chapter 5 1 3 Please note that when using an Incubation Chamber 4 Slides product numbers 12...

Page 23: ...gether with the ibidi Heating System Slide Dish Silver Line The Gas Mixer and the Humidifying Column should be placed next to the microscope and on the same height level as the microscope stand The tubing of the Heated Tubing Package needs to reach the Incubation Chamber on the microscope stage ibidi Gas Mixer ibidi Temp Controller Heated Tubing Package Electrical cable Gas tubing Computer optiona...

Page 24: ... supply Type Serial number Production date ibidi Gas Mixer ibiGM2 001 11 2018 ibidi GmbH Am Klopferspitz 19 D 82152 Martinsried Germany 49 0 89520 46 17 0 www ibidi com Output 1 Input N 2 Input Air Input CO 2 Output 2 Figure 10 Overview of the connection of all cables and tubing to the Gas Mixer In the CO2 version the N2 port is not available as closed by a seal 5 1 1 Connection to the Gas Supply ...

Page 25: ...ect B CAUTION Always connect all gas input ports with the appropriate gas 1 bar 14 5 psi Do not leave any input ports open CO2 O2 version In case the N2 input is not needed CO2 O2 version only close the N2 input port using the provided dead end tubing Figure 13 N2 Position OFF Figure 13 Close the N2 port of the Gas Mixer with the dead end tubing if not needed CO2 O2 version only To connect the gas...

Page 26: ...e Humidifying Column the other directly to the Incubation Chamber Table 6 Connection of the gas lines with color code of the mixed gas Color Code Gas Mixer output 1 white Blue tubing with white label Gas Mixer output 2 yellow Blue tubing with yellow label Humidifying Column output red Blue tubing with red label Humidifying Column output blue Blue tubing with blue label 1 Connect the two gas outlet...

Page 27: ... Serial number Production date ibidi Gas Mixer ibiGM2 001 11 2018 ibidi GmbH Am Klopferspitz 19 D 82152 Martinsried Germany 49 0 89520 46 17 0 www ibidi com max 50 C 122 F CAUTION HOT SURFACE Incubation Chamber Slide Dish Push in fitting Figure 15 Connection of the Heated Tubing Package to the Gas Incubation System CAUTION Do not disconnect the Heated Tubing while the Gas Mixer is running The resu...

Page 28: ...ion date ibidi Gas Mixer ibiGM2 001 11 2018 ibidi GmbH Am Klopferspitz 19 D 82152 Martinsried Germany 49 0 89520 46 17 0 www ibidi com 1 2 Type Serial number Production date ibidi Temperature Controller ibiTC3 001 11 2021 ibidi GmbH Am Klopferspitz 19 D 82152 Martinsried Germany 49 0 89520 46 17 0 www ibidi com Temp Sensor Power Signal Out 3 6 4 5 Output Channels 24 V max 6 7 A USB On Off Gas Mixe...

Page 29: ... 5 2 Re Filling the Humidifying Column Before the first use the Humidifying Column must be filled with 130 ml of ultra pure water During operation the column should always be refilled before the water level falls below the minimum mark max 50 C 122 F CAUTION HOT SURFACE Water Level max 130 ml Water Level min 50 ml Figure 17 Re Filling the Humidifying Column Open the blue stopper on top of the colu...

Page 30: ...on the display Section 5 4 4 Check that the display shows Run see Section 19 5 Let the system equilibrate all parameters for a minimum of 30 minutes NOTE Immediately after powering up the Gas Mixer runs a flow sensor calibration routine for several seconds The display shows self test After this the Gas Mixer for CO2 O2 will automatically start a warm up routine for 5 minutes The display shows a bl...

Page 31: ...he chan nels The cursor position is indicated with square brackets You can move the cursor using the left and right buttons If you want to select a parameter or a function press the round button and the square brackets will change to angle brackets Now you are able to change the value with the up and down buttons To confirm the changed value you must press the round button once more 5 4 1 Run Stop...

Page 32: ...and the temperature of the measured environment Flow rate At flow rates below 7 l h gas mixes may not reach the highest possible humidity If this is the case augment the gas flow rate until the humidity is reached O2 regulation hypoxia Gas Mixer for CO2 and O2 only To generate hypoxia conditions set the oxygen to the target value The O2 will be replaced by nitrogen N2 until the set value is reache...

Page 33: ...l of the Gas Mixer can be set to an On or Off state For standard operation all channels must be turned on Only if you use the CO2 O2 version without nitrogen N2 connection it is recommended to turn the O2 channel off In this case make sure that the dead end tubing is attached to the N2 port and closed Mode Setup Channel 1 Mode On Return Figure 21 Mode dialog 1 Press the round button once more to m...

Page 34: ...h individual channel If the alarm is activated which happens when the current value goes under over the low high limits the current value I blinks showing alternately the value and low or high respectively To stop the display blinking navigate to the blinking channel and press the round button once Alarms Setup Channel 4 High 42 C Low 36 C Figure 22 Alarm settings dialog Preferences Set the bright...

Page 35: ... ibidi website For more details please refer to the IncuControl instructions Figure 25 Gas Mixer control window in IncuControl Prohibitions on the use of ibidi software The following actions are prohibited Copying software for other than backup purposes Transfering or licensing of the right to use software to a third party Disclosure of confidential information regarding software Modification of s...

Page 36: ...he Humidifying Column is usually not a problem since distilled water and light starved environments microscopy rooms do not support microbe growth If however contamination has occurred disinfect the Humidifying Column by running the system with 3 hy drogen peroxide for 1 hour Subsequently wash with water The use of anti microbial agents is not recommended since they can be transported as aerosols ...

Page 37: ... Con nected in the IncuControl software or does not appear on the device display Gas mixer for CO2 only Make sure the Gas Mixer is a CO2 O2 mixer CO2 only mixers do not have an O2 channel The CO2 or O2 gas channel displays large negative values or the error mes sage SHORT is displayed Contact ibidi technical sup port 8 2 Flow Readings Error Description Possible Cause Possible Solution Gas flow rea...

Page 38: ...e dead end tub ing and set the O2 channel OFF on the de vice display or IncuControl software Defective valve or flow sensor Turn off the gas mixer and check if gas is still flowing through the Humidifying Col umn appearance of gas bubbles inside If gas is still flowing inside the Humidifying Column the valve is defective Contact ibidi technical support Gas line restriction Make sure that the gas l...

Page 39: ...Defective valve or flow sensor Contact ibidi technical support CO2 regulation is very slow CO2 level has not reached the set value 2 Let CO2 level adjust for 15 minutes CO2 The channel has been turned ON just after it was OFF for a while It will take up to 10 minutes for the CO2 level to reach the adjusted set point 3 8 4 O2 Channel Readings Error Description Possible Cause Possible Solution O2 ch...

Page 40: ...ing gas is not connected Additionally keep in mind that the maximum O2 concentration depends on the set CO2 concentration See chapter 5 4 2 for further details 8 5 Humidity Channel Readings Error Description Possible Cause Possible Solution Humidity channel reading is 0 Humidity sensor not con nected Connect the Incubation Chamber as shown in Figure 16 Condensation on the humid ity sensor Open and...

Page 41: ...s seen Contact ibidi technical sup port Humidifying Column not closed properly Make sure that the column is properly sealed with the blue silicone plug Defective Humidifying Col umn Check if the Humidifying Column is heat ing up and measure the temperature of the water inside the Humidifying Column set point temperature must be 45 50 C Check IncuControl or the display to see if the set temperature...

Page 42: ...on time lapse ex periments Focus stability is mainly influenced by mechanical changes and temperature variations Follow these recommendations to keep your cells in focus Switch on all components e g heating gas incubation computer or other equipment at least 60 minutes before starting the experiment After you put the sample onto the microscope wait 30 minutes before starting a time lapse experimen...

Page 43: ... inside the incubator overnight This step helps to avoid the formation of air bubbles and pre warms the solutions to 37 C Afterwards fill your slide with cells and medium Cover the medium s surface with an appropriate amount of silicone oil Don t drip the oil directly onto the surface but let it run down the edges of the cell culture vessel Details about avoiding evaporation are given on the ibidi...

Page 44: ...chhamer Schlag 11 82166 Gräfelfing Germany Toll free within Germany Phone 0800 00 11 11 28 Fax 0800 00 11 11 29 International calls Phone 49 89 520 46 17 0 Fax 49 89 520 46 17 59 E Mail info ibidi com ibidi com com ...

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