Instruction Manual for the ibidi Heating System 1 ... · Instruction Manual for the ibidi Heating System 1, Universal Fit Version 2.1 ... variety of optical microscope setup requirements

Instruction Manual for the

ibidi Heating System 1, Universal Fit

Version 2.1

ibidi Heating System 1, Instruction Manual

II ©ibidi GmbH 2012, version 2.1, revision 1, 2012-12-03, Dr. Miro Kroutvar, Dr. Zeno Guttenberg


© ibidi GmbH 2012, version 2.1, revision 1, 2012-10-09, Dr. Miro Kroutvar, Dr. Zeno Guttenberg 1

Dear User,

Live cell imaging enhances the power of cell microscopy, immensely. Intracellular transport

phenomena can be tracked with high-resolution fluorescence microscopy. In addition, the

low-magnification microscopy of living cells, such as is used for migration or proliferation

experiments, is a topic of great interest. From these examples, you can appreciate the wide

variety of optical microscope setup requirements that are often used in one lab.

So, when considering a design for the ibidi Heating System, our objective was to create a

microenvironment for cell culture and microscopy that would be compatible with every type

of inverted microscope, regardless of the brand.

We then created the following list of requirements for the new heating system, based on

customer needs and the industry standards:

• Stable conditions for temperature, humidity, CO2, and O2

• Compatible with any inverted microscope

• Suitable for, but not limited to, use with all ibidi µ-Slides and µ-Dishes

• Usable with all objective lenses (up to 100x oil)

• Portable from one microscope to another

• Easy handling

• Reasonable pricing

• Short installation time

• Reversible; no modification to your microscope required

• Usable within an incubated microscope

After considering these issues, we came up with the idea to search for a standard format

that could be used on all inverted microscopes. Our solution was simple: a multi-well format.

There are a large numbers of plates currently in use (e.g., 12 well, 24 well, 96 well, etc.) and

all of them have the same outer dimensions: 127.5 mm x 85.5 mm. This means that for

every inverted microscope, there is a stage or a holder in the same format. This type of

holderbecame the basis for all of the microscope platforms.

Using this multi-well idea, we designed a heating system that is capable of meeting all of

our requirements. Universal usage is just one of the system’s outstanding features. We also

developed a heated lid that prevents water condensation effects and provides stable

temperature conditions. Finally, controls for CO2, O2, and humidity complete the system.

Thank you for purchasing the ibidi Heating System. We wish you success with it, and we

welcome your feedback!


2 ©ibidi GmbH 2012, version 2.1, revision 1, 2012-12-03, Dr. Miro Kroutvar, Dr. Zeno Guttenberg

Table of contents

Safety Considerations............................................................................................................ 4

Regulatory Statement............................................................................................................ 7

Specifications ........................................................................................................................ 8

Preface .................................................................................................................................10

Notices .................................................................................................................................10

Limited Warranty ..................................................................................................................11

Installation Requirements .....................................................................................................12

Installation and Connecting Cables .......................................................................................13

Maintenance and Operation .................................................................................................14

Waste Treatment .................................................................................................................15

1 Working Principle ..........................................................................................................16

2 Equipment .....................................................................................................................17

2.1 Product Description ..............................................................................................17

2.2 Heated Lid ............................................................................................................17

2.3 Heated Plate and Inserts .......................................................................................18

2.4 Sterilization and Cleaning ......................................................................................19

3 ibidi Temperature Controller ..........................................................................................20

3.1 Connecting the Temperature Controller ................................................................21

3.2 Menu Navigation ...................................................................................................22

3.3 Supply Voltage Adjustments .................................................................................22

3.4 Setting into Operation ...........................................................................................22

4 Temperature Calibration ................................................................................................24

4.1 Calibration Routine ................................................................................................24

4.2 Influence of Surrounding Temperature and Airflow ...............................................25

5 TempControl Software ..................................................................................................27

5.1 Software Overview ...............................................................................................27

5.2 Logging Menu .......................................................................................................29

5.3 Advanced Menu ....................................................................................................29

5.3.1 Calibration Routine Using TempControl .............................................................30

6 Troubleshooting ............................................................................................................31

6.1 Focus Not Stable ...................................................................................................31

6.2 Evaporation Too High ............................................................................................31

6.2.1 Parafilm Procedure ............................................................................................31

6.2.2 Silicone Oil Procedure (ibidi’s Anti-Evaporation Oil) ...........................................31

7 WEEE/RoHS Compliance Statement .............................................................................33

A Menu Navigation ............................................................................................................. I

A.1 Setting the Temperature ......................................................................................... I

A.2 External Temperature Sensor ................................................................................. II



A.3 Run / Stop Button .................................................................................................. II

A.4 Setup Button ..........................................................................................................III

A.4.1 Mode Function ...................................................................................................III

A.4.2 Alarm Function .................................................................................................. IV

A.4.3 Preferences ........................................................................................................ V

A.4.4 Calibration .......................................................................................................... V

A.4.5 Info ................................................................................................................... VI



Safety Considerations

To ensure operation safety, the ibidi heating system must be operated correctly and

maintained according to a regular schedule. Carefully read to fully understand all safety

precautions in this manual before operating the instrument. Please take a moment to

understand what the signal words WARNING!, CAUTION and NOTE mean in this manual.

Safety symbols

WARNING! A WARNING! indicates a potentially hazardous situation which,

if not avoided, could result in serious injury or even death.

CAUTION A CAUTION indicates a potentially hazardous situation which, if

not avoided, may result in minor or moderate injury. It may also

be used alert against damaging the equipment or the

instrument.

Do not proceed beyond a WARNING! or CAUTION notice until

you understand the hazardous conditions and have taken the

appropriate steps.

NOTE A NOTE provides additional information to help the operator

achieve optimal instrument and assay performance.

Read manual label. This label indicates that you have to read

the manual before using the instrument. This label is positioned

at the backside of the temperature controller.

This symbol indicates important notes.

Identification label. This label is positioned at the

backside / rear panel of the temperature controller.



Nomenclature

TEMPERATURE HEATED LID HEATED PLATE

CONTROLLER

WARNING! Do only operate the ibidi heating system with the delivered cables and plugs. If

not doing so you risk electric shock and fire.

WARNING! Do not operate the ibidi heating system with substances and under conditions

which do cause a risk of explosion, implosion or release of gases. Only operate the ibidi

heating system with aqueous solutions.

CAUTION Ensure that the power plug is well accessible. The ibidi heating system has to be

installed in a way that it does not hinder the access to the power plug.

CAUTION Operate the heating system only on a power socket with protective conductor to

ground the housing of the temperature controller.

CAUTION The weight of the ibidi heating controller is approx. 3.4 kg. Provide a secure stand

for controller, where it cannot be accidentally dropped. Moving the heating system poses a

risk of personal injury or damage to the instrument.

CAUTION The manual opening of the temperature controller is not allowed. Manual opening

pose a risk of personal injury or damage to the controller. Contact ibidi service personnel if

you need to open the controller.

CAUTION The ibidi heating system includes a heated plate with heated lid to control the

sample temperatures. Some accessible parts of the plate and lid can reach temperatures up

to 55°C. Avoid touching the temperature controlled parts of the system when you have set

the temperature controller to high temperatures.



CAUTION When the glass plate of the heated lid was broken due to mechanical impact, the

shards can lead to injuries if touched.

CAUTION Be aware that when the system is switched on, 10 V AC voltage is applied to the

underside of the glass plate. Do not touch the underside or contact it with anything

conductive. This could cause a short circuit that may destroy the temperature controller

and/or the lid.

CAUTION Only use correct replacement fuses. The correct values of the replacement fuses

are indicated on the backside of the heating controller. Using wrong replacement fuses can

lead to overheating of the temperature controller which can cause fires.

CAUTION Only ibidi staff is allowed for servicing and opening of the temperature controller.

Turn off the power switch and unplug the power cord before servicing the system, unless

otherwise noted. Connect the temperature controller only to the delivered power plug. Do

not use extension cords. Have an electrician immediately replace any damaged cords, plugs,

or cables. Not doing so poses a risk of personal injury or damage to the system.

CAUTION Do only use the heating system in dry rooms (and do not use the heating system

in the cold room.

CAUTION Turn off the power switch in the back of the chassis, when the heating system is

not in use.

CAUTION Do not use ethanol or other types of organic solvents to clean the heating system

as they may remove the paint.

CAUTION Use the instrument only for cell experiments with aqueous solutions.

CAUTION Do not use the instrument with hazardous substances or substances/materials

which pose a risk of infections.



Regulatory Statement

in preparation

The following safety and electromagnetic standards were considered:

• IEC 61010-1:2001 Safety requirements for electrical equipment for measurement,

control and laboratory use. Part 1 General Requirements

• IEC 61010-2-010:2005 Safety requirements for electrical equipment for

measurement, control and laboratory use. Part 2-010: Particular requirements for

laboratory equipment for the heating of materials.

• IEC 61010-2-081:2001 Safety requirements for electrical equipment for

measurement, control and laboratory use. Part 2-081: Particular requirements for

automatic and semi-automatic laboratory equipment for analysis and other purposes.

• IEC 61326-1:2006 EMC, Electrical equipment for measurement, control and

laboratory use – EMC requirements.

• IEC 61000-3-2:2006 EMC, Limits for harmonic current emissions (equipment input

current up to and including 16A per phase).

• IEC 61000-3-3:2008 EMC, Limits



Specifications Heating System 1

Power Consumption

Supply voltage 230 V / 50 Hz - 115 V / 60 Hz

Output voltage 10 – 40 V

Max. power input 160 VA

Max. power output 160 VA

Surrounding Conditions

Storage Temperature -5 – 50°C

Operating Temperature 15 - 40°C (Indoor use only)

Humidity 80 % relative humidity up to 31°C,

30 % relative humidity up to 40°C

Operating altitude max. 2000 m

Temperature Control Range Ambient temperature to +55°C

Net Dimensions (hxwxd)

Controller 90 x 170 x 230 mm³

Heated Plate with Lid 25.5 x 125.5 x 85.5 mm³

Weight

Controller 3.4 kg

Heated Plate with Lid 210 g

Fuses DIN 41661 (5 x 20 mm)

230 V / 50 Hz 1 A (T)

115 V / 60 Hz 2 A (T)



Connections to the temperature controller

All ingoing and outgoing connections can be found at the rear panel of the instrument.

Name Function

USB cable

Power Switch

Power cable

Fuse box

External sensors

Control channels 1-4

Socket for connecting the USB cable to the PC/Notebook. To

setup a computer communication to the ibidi temperature

controller, the USB cable must be connected.

When the switch is pressed on (position “I”), the temperature

controller is switched on.

The connector to the electrical power supply

Contains the fuses for the temperature controller

Ports for external temperature sensors (four terminal

sensing of Pt100 resistors and thermocouple sensors)

4 connectors for the 4 control channels



Preface

This manual is your guide for using the ibidi heating system for cell culture experiments on

an optical microscope. It instructs first-time users on how to use the instrument, and serves

as a reference for experienced users.

Before using the ibidi heating system, please read this instruction manual carefully, and

make sure that the contents are fully understood. This manual should be easily accessible to

the operator at all times during instrument operation. When not using the instrument, keep

this manual in a safe place. If this manual becomes lost, order a replacement from ibidi

GmbH.

Notices

1. ibidi shall not be held liable, either directly or indirectly, for any consequential damage

incurred as a result of product use.

2. Prohibitions on the use of ibidi software

• Copying software for other than backup

• Transfer or licensing of the right to use software to a third party

• Disclosure of confidential information regarding software

• Modification of software

• Use of software on multiple workstations, network terminals, or by other

methods

3. The contents of this manual are subject to change without notice for product

improvement.

4. This manual is considered complete and accurate at publication.

5. This manual does not guarantee the validity of any patent rights or other rights.

6. If an ibidi software program has failed causing an error or improper operation, this

may be caused by a conflict from another program operating on the notebook (PC). In

this case, take corrective action by uninstalling the conflicting product(s).

7. ibidi is a registered trademark of ibidi GmbH in Germany and other countries.



Limited Warranty

Products sold by ibidi, unless otherwise specified, are warranted for a period of one year

from the date of shipment to be free of defects in materials and workmanship. If any

defects in the product are found during this warranty period, ibidi will repair or replace the

defective part(s) or product free of charge.

THIS WARRANTY DOES NOT APPLY TO DEFECTS RESULTING FROM THE FOLLOWING:

1. IMPROPER OR INADEQUATE INSTALLATION.

2. IMPROPER OR INADEQUATE OPERATION, MAINTENANCE, ADJUSTMENT OR

CALIBRATION.

3. UNAUTHORIZED MODIFICATION OR MISUSE.

4. USE OF UNAUTHORIZED CELL CULTURE WARE.

5. USE OF CONSUMABLES, DISPOSABLES AND PARTS NOT SUPPLIED BY AN

AUTHORIZED IBIDI DISTRIBUTOR.

6. CORROSION DUE TO THE USE OF IMPROPER SOLVENTS, SAMPLES, OR DUE TO

SURROUNDING GASES.

7. ACCIDENTS BEYOND IBIDI’S CONTROL, INCLUDING NATURAL DISASTERS.

This warranty does not cover consumables like cell culture dishes, reagents and the like.

The warranty for all parts supplied and repairs provided under this warranty expires on the

warranty expiration date of the original product. For inquiries concerning repair service,

contact ibidi after confirming the model name and serial number of your ibidi system.



Installation Requirements

To ensure operation safety, observe the following conditions:

1. Do only operate the ibidi heating system with the delivered power cable.

2. Only connect the power cable of the ibidi heating system to an electrical socket

containing a protective conductor terminal.

3. Ensure that the power plug of the power supply is well accessible. The ibidi heating

system has to be installed in a way that it does not hinder the access to the external power

supply and its power plug.

4. Only operate the heating system with the delivered PC (Notebook).

5. Only operate the heating system with original ibidi insert.

6. Operate the heating system in a temperature range of 15 – 40°C.

7. Operate the heating system in a humidity range of 0 – 80% RH up to 31°C and 30% RH

up to 40 °C.

8. Operate the heating system in an atmospheric pressure range of 800 – 1060 hPa.

9. Do not operate the heating system under conditions which pose a risk of explosion,

implosion or the risk of the release of gases.

10. Avoid strong magnetic fields and sources of high frequency. The heating system may

not function properly when near a strong magnetic field or high frequency source.

11. Avoid vibrations from vacuum pumps, centrifuges, electric motors, processing

equipment and machine tools.

12. Avoid dust and corrosive gas. Do not install the heating system where it may by exposed

to dust, especially in locations exposed to outside air or ventilation outlets.

13. For cleaning the heating system, only use ultrapure water.

14. Do not install the heating system in a location where it may be exposed to direct

sunlight.

15. Install the heating system in a horizontal and stable position. This includes a table, bench

or desk upon which the instrument is installed.

16. Ensure that no air conditioner blows air directly onto the heating system. This may

prevent stable conditions.

17. Install the heating system in a location that allows easy access for maintenance.

NOTE: The above conditions do not guarantee optimal performance of this heating system.



Installation and Connecting Cables

Connecting the ibidi heating system to the power supply

Confirm that the power switch of the heating system is off (power switch is at the backside,

right of the temperature controller).

WARNING! Do only operate the ibidi heating system with the delivered cables and plugs. If

not doing so you risk electric shock and fire.

CAUTION Ensure that the power plug of the external power supply is well accessible. The

heating system has to be installed in a way that it does not hinder the access to its power

plug.

Connect the heating system to the Notebook by using the delivered network cable.

Switch on the heating system and the Notebook.



Maintenance and Operation

Pay attention to the heating system operating environment and always keep it clean so that

the instrument can be used in a stabilized condition over a long period. Do not place

anything heavy on the temperature controller.

Cleaning the ibidi heating system

Switch off the heating system and remove the power plug from the electrical socket. Only

use dry cloth or cloth wetted with water (ultrapure) for cleaning of the instrument.

CAUTION Do not use ethanol or other types of organic solvents to clean the instrument as

they may remove the instrument paint.

Transporting the ibidi heating system

Switch off the temperature controller and remove the power plug from the electrical socket.

Carry the instrument carefully and avoid mechanical shocks.

CAUTION The weight of the temperature controller is approx. 3.4 kg. Dropping the

instrument while moving it, may cause personal injury or damage.

Functional Disorder

In case of a functional disorder switch off the temperature controller and wait for five

minutes, then switch on the instrument again. If there is still a functional disorder switch off

the device, unplug the power cable and contact the ibidi service.

Repairing the ibidi heating system

Do not try to repair the instrument by yourself. Do contact the ibidi service for repairing the

instrument.

CAUTION The manual opening of the instrument is not allowed. Manual opening pose a risk

of personal injury or damage to the instrument. Contact ibidi service personnel if you need to

open the instrument.



Waste Treatment

Waste treatment is your own responsibility. You must hand it to a company specialized in

waste recovery. Do not dispose the ibidi heating system in a litter bin or at a public waste

disposable site. For detailed information please contact the ibidi service.



1 Working Principle

The stage top incubator consists of a heated plate, and also a heated lid that is designed to

keep cells at 37°C in microscopy slides and dishes. The heated lid prevents condensation

effects inside the entire incubation system. Humidity, CO2 enriched, and/or O2

reduced/enriched air can be flushed in, which makes the heated chamber a fully controlled

incubator—right on the microscope. This enables the user to perform live cell imaging.

The glass top of the lid and the bottom plate are actively heat-controlled. The heated plate

has the outer dimensions of a 96-well plate (127.5 mm x 85.5 mm).

Figure 2: Heated Lid and Heated Plate (without Insert)

Figure 1: Schematic Drawing of the ibidi Heating System



2 Equipment

Due to the height of the lid, we recommend using condensers with a working distance of

≥ 26 mm.

Information on gas incubation (CO2, O2, humidity) is provided in the Add-On instructions:

“Gas Incubation with Active Gas Mixer”.

2.1 Product Description

A brief overview is provided in this section, including the nomenclature for later descriptions

in this document.

Figure 3: Schematic Setup

2.2 Heated Lid

The heated lid (127.5 mm x 85.5 mm) provides excellent optical quality, and also allows for

the use of all standard microscopy techniques, including differential interference contrast

(DIC). The upper glass part is electrically heat controlled. The heated lid fits accurately to the

heated plate.

For gas incubation, the lid is equipped with inlets for (a) the gas flow and (b) the humidity

sensor.

Optional: PC

Figure 4: Heated Lid

Temperature

controller

Incubation

chamber



2.3 Heated Plate and Inserts

The heated plate provides the base for the heating system. Its format (127.5 mm x 85.5

mm) fits every multi-well stage or multi-well holder (e.g., for 96-well plates) on the

microscope. It is fully heated with electricity. The heated plate is the counterpart to the

heated lid.

The two-part insert has two functions that both use the force of the integrated magnets. The

first function is to hold the µ-Slides or µ-Dishes firmly into position, so as to avoid

displacement during microscope stage movements. The second is to create a tight contact

of the µ-Slides or µ-Dishes to the metallic insert, which then maximizes the heat transfer and

ensures the stable heat control of the sample. The heated plate passively heats the inserts.

Figure 5: Heated Plate with Inserts

Heated plate, heated lid, and inserts are positioned by strong Neodym

Magnets! Please contact us for a non-magnetic system, in the case

that permanent magnetic fields will disturb your experiment!



2.4 Sterilization and Cleaning

The heated plate, the heated lid, and the inserts do not have to be sterile, because the

incubation chamber has no direct contact with the cells and the cell media.

However, all these parts can be cleaned. The inserts are very robust and can be cleaned

with Ethanol. The lid is made of polycarbonate and, hence, has only limited resistance

against alcohol. Therefore, we recommend using ultrapure water for cleaning. Also, the

bottom of the heated plate is not alcohol resistant, so this plate should not be cleaned with

Ethanol. Please use only the ultrapure water.

Do not clean lid and plate with alcohol! Use ultrapure water

(e.g., Millipore water) instead!



3 ibidi Temperature Controller

The ibidi Temperature Controller is designed to precisely regulate the ibidi incubation

chamber. To achieve this, the controller has four control channels on the rear panel. When

required, the controller can be customized for two, or up to 64 channels. In addition, this unit

can be run as a standalone system or PC-controlled.

In Figure 6, you can see a picture of the controller. The measured (‘I’) and set (‘S’)

temperatures of all the channels are displayed. You can navigate through the menu by using

the control buttons on the right. The LEDs, located below the display, indicate the status of

the channels and feedback, the power, and the USB connection.

Figure 6: Front Panel of the ibidi Temperature Controller

Up /

rightbutton

Select /

confirmbutton

Down /

leftbutton

Control LEDs

LC display

Figure 6: Front Panel of the ibidi Temperature Controller

Up /

Rightbutton

Select /

Confirmbutton

Down /

Leftbutton

Control LEDs

LC display



3.1 Connecting the Temperature Controller

All plugs for the electrical connections are integrated into the rear of the ibidi Temperature

Controller, as shown in Figure . On the right side, you will find the plug for the power cable.

Above that is the power switch, and the fuse box is at the top. The USB cable connection is

located in the center of the box. There are ports for external temperature sensors (four-

terminal sensing of Pt100 resistors and thermocouple sensors). On the left side, you will

find the four connectors for the four control channels. In addition to these plugs, there are

upgrade options which you can find below.

The standard connectors are:

• power cable

• USB cable

• 4 control channels

• 2 external temperature sensors

Optional features:

• USB stick

• customized connector to for up to 64 control channels

• reduction to 2 control channels

To setup a computer communication to the ibidi Temperature Controller, the USB cable

must be connected.

Figure 7: Rear Panel of the ibidi Temperature Controller

Power

cable

Power

switch

Fusebox

USB cable

Controlchan

nels 1-4

Externals

ensors



To calibrate the system for the sample temperature, you can use the external temperature

sensor of the controller or the control software TempControl. For more details, please refer

to Section 4.

3.2 Menu Navigation

Most control parameters can be manually set on the controller using the buttons and the LC

display on the front panel. The functions of the button are:

• up / right

• down / left

• select / confirm

The cursor position is indicated with square brackets (‘[xxx]’). You can move the cursor using

the ‘Left’ and ‘Right’ buttons. If you want to select a parameter or a function, you need to

press the ‘Select’ button. When you select a parameter, the square brackets will change to

angle brackets (‘<xxx>’). Now you are able to change the value up or down. To confirm the

changed value, you must press the ‘Confirm’ button once more. When selecting a function,

you will see the individual function view, which is where you can change the parameters or

access further functions.

You can find a detailed description of the menu navigation, including all functions, in

Appendix A.

3.3 Supply Voltage Adjustments

The control unit should only be used in dry rooms.

The control units must be adjusted to the corresponding voltage (230 V or 115 V). The actual

voltage setting may be read and adjusted at the integrated fuse box of the power cable

connector, located at the rear panel. In order to change these settings, it is necessary to pull

out the fuse holder from the casing. Replace the fuses to match the input voltage (1 A(T) for

230V / 50Hz or 2 A (T) for 115 V / 60 Hz). Rotate the grey fuse holder, and then reinsert it so

that the correct voltage is shown in the window. Snap the fuse box back into the housing.

The ‘On/Off’ switch is at the rear side of the unit. After selecting ‘On’, the controller will

automatically start a self-test, and then the measured values will be shown.

3.4 Setting into Operation

Here is the procedure to start the operation of the controller.

Be aware that when the system is switched on, 10V AC

voltage is applied to the underside of the glass plate. Do not

touch the underside or contact it with anything conductive.

This could cause a short circuit that may destroy the

controller and/or the lid.



For the first power up, we recommend disconnecting the heating devices (e.g., the heated

lid and the heated plate). By doing this, you ensure that you do not start heating with the

wrong temperature settings. It is now possible to set the temperatures for the individual

channels. Afterwards, you can reconnect the heating devices.

Since the controller is preconfigured, we recommend connecting the heated lid to Channel 1

and the heated plate to Channel 2. For subsequent power ups, you will not need to

disconnect the heating devices.

Please let the temperature of the system equilibrate for a

minimum of 30 minutes before you start your experiments.



4 Temperature Calibration

Depending on the room temperature, air conditioning, heat loss via microscope, chamber

type or objective lens, and humidification, the temperature at the position of the cells has to

be calibrated from time to time. A calibration is recommended before beginning the first

experiment. The heat calibration should be done for each chamber type, in order to control

temperatures with an absolute accuracy of less than 1 °C.

For calibration, the ibidi Temperature Controller has a plug, located on the rear panel, for an

external temperature sensor. Before shipment, all controllers are run through an in-house

calibration to ensure their accuracy. This calibration only refers to the actively heated

devices, such as the plate and the lid. The samples are only indirectly heated via the insert.

As a result, the cell temperature will lie somewhat below the temperature of the heated

plate. To compensate for this effect, the heated plate can be calibrated to the cell

temperature that is sensed by the external temperature sensor, which can be placed at the

position of the cells.

4.1 Calibration Routine

The recommended calibration routine is fairly simple. Follow these eight steps for the initial

temperature calibration:

1) Plug the external temperature (thermo-couple type K) sensor into the rear panel of

the temperature controller. Select the correct input for display of the sensor (TC-K,

see Section A.2).

2) Fill the µ-Slide or µ-Dish with medium and place the sensor as close as possible to

the later position of the cells. Use the prepared calibration set, which consists of a µ-

Dish or a µ-Slide with a perforated lid, as shown in the Figure .

3) Set all parameters to create conditions identical to those of the actual experiments,

such as room temperature, air conditioning, airflow, illumination, microscope settings,

etc.

4) Set the temperature of heated plate to the desired cell temperature and set the

temperature of the lid several degrees higher (e.g., plate 37 °C, lid 42 °C). You can do

this directly with the control buttons on the controller, or with the TempControl

software.

5) Let the system equilibrate for 30 minutes.

6) Go to the ‘Calibration’ mode of the controller (see Section A.4.4) and execute the

‘Calibrate Now’ function for the channel called ‘Plate’, because the heated plate is the

main heat source for the cells.

The displayed temperatures are valid for the heated

plate and the heated lid only. The temperature near

the cells might be different (lower in most cases)!



7) If you use the TempControl software, you can calculate the difference between the

measured cell temperature with the external sensor and the displayed temperature.

This value can then typed in as the ‘Offset’ in the corresponding window of the

channel configuration. This method is described in more detail in Section 5.3.1.

8) Check the temperature of the system from time to time.

Figure 8: Sensor Placement for Temperature Calibration

4.2 Influence of Surrounding Temperature and Airflow

The surrounding, ambient temperature affects the temperature inside the microscopy

chambers. Devices, such as computers and camera controllers, can significantly heat up

small rooms. In this case, we recommend equilibrating the room temperature to the typical

experimental conditions at least 2-3 hours before calibrating.

Keep in mind that the temperature sensor can also measure room temperatures.

You will be able to calibrate any other connected, heated

device in the same way.



The airflow can enhance the effect of temperature changes in the vicinity of the incubation

chamber. In a case where the airflow (e.g., air conditioning, etc.) cannot be stopped, we

recommend protecting the microscope as much as possible from any strong airflow. Please

contact us for support.



5 TempControl Software

The ibidi Temperature Controller has an USB interface for computer control. The

TempControl software is the tool that was developed for this remote control, and comes

with the controller.

5.1 Software Overview

In the main view window of the TempControl software, you can set the temperature for

each individual channel, plus you can view the measured temperature, and also the

temperature course. Additionally, you can find indicators for the channel status and the

communication between the controller and the computer.

From this view, you can access further sub-menus. By

activating the ‘Monitoring’ button, you get a minimized view of

the measured temperatures for each channel, as shown in

Figure . You can use this feature when your desktop space is

limited.

By clicking inside the temperature graph, you can expand the

graphical presentation of the temperature course, as seen in

Figure .

Figure 9: Main View of TempControl Window

Graphical temperature progress

Click to expand

Activate minimized

temperature monitor

Channel status

Channel ON/OFF switch

Measured temperature

Set temperature

Figure 10: Minimized View

of the Temperature

Monitoring Window



The ‘Logging Menu’ button brings you to a menu where you can start logging in the

temperature course. The ‘Advanced’ button leads you to a menu where the channel

parameters can be viewed and modified.

Figure 11: Graphical Presentation of the Temperature Course



5.2 Logging Menu

If you choose ‘Start Logging’, you have to select a file name under which the data should be

stored. This data can be viewed in a normal text viewer or it can be loaded using the ‘Open’

button.

There are two further options for the data logging. If you select ‘Advanced Logging’, you can

save disk space, because only the last 30 minutes will be stored. If you select the ‘Append

Values’ box, then, in addition to the last 30 minutes, all data after an incident is recorded, as

in the case of a temperature alert.

5.3 Advanced Menu

The ‘Advanced’ menu allows you to configure the individual channel parameters, starting

from the PID algorithm coefficients over the input limits, to the alarm settings. There, you

can also label the channels according to the heating device. You will find that Channel 1 is

labeled ‘Lid’ and Channel 2 is labeled ‘Plate’.



An interesting option for many users is the possibility of defining a temperature offset. This

is a simple way to calibrate the controller.

5.3.1 Calibration Routine Using TempControl

Using the external temperature sensor, measure the temperature at the cell position, as is

shown in Section 4.1. Calculate the difference between the temperature measured by the

integrated sensor of the heating device and that of the external sensor. Use this value as the

offset. If, after changing the offset the temperature still deviates, repeat this step.

Figure 12: Advanced Menu for Control Channel Configuration



6 Troubleshooting

6.1 Focus Not Stable

Focus drift is a disturbing effect, especially during time-lapse experiments. Focus stability is

mainly influenced by mechanical changes and temperature variations. Follow these

recommendations to keep your cells in focus:

• Switch on all components (heating, gas incubation, computer, other equipment) at

least 60 minutes before starting the experiment.

• After you put the µ-Slide onto the microscope, wait 20 minutes before starting a

time-lapse experiment to achieve temperature and immersion oil equilibration. (*)

• Keep the room temperature as stable as possible. Air conditioning should either be

working continuously or switched off.

• Do not change the temperature during the experiments. Avoid door/window

openings, as this could rapidly change the temperature.

• Eliminate all sources of mechanical vibrations. Use a damped table for your

microscope.

(*) In case the experiment needs to be started immediately, either after placing the slide on

the microscope, or after closing the lid, we recommend controlling the focus for 20 minutes.

In the first minutes after starting the experiment, temperature equilibration might influence

the focus/z-position of the cells.

6.2 Evaporation Too High

Depending on the incubating conditions, small volumes might evaporate quickly, especially

during long-term experiments. We suggest using Parafilm (Pechiney Plastic Packaging

Company) or silicone oil AR 200 (Fluka, Sigma-Aldrich) to decrease evaporation. If you have

an actively controlled humidifying device, increase the set value of relative humidity.

6.2.1 Parafilm Procedure

Small stripes of Parafilm fitted onto the reservoirs of the slides are very effective at

preventing evaporation effects. Fill the slide, as recommended, then adapt the Parafilm until

it fits tightly.

6.2.2 Silicone Oil Procedure (ibidi’s Anti-Evaporation Oil)

Covering the medium with sterile silicone oil prevents all evaporation effects and is

compatible with cell culture. Please don’t use mineral oil, as this would be harmful to the

ibidi µ-Slides.

Equilibrate oil and medium inside the incubator overnight. This step helps 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 by pressing the



pipette tip directly on the upper side of the reservoir. For example, when using µ-Slide

VI 0.4, fill each reservoir with 30 µl cell-free medium and 30 µl silicone oil.

Please contact us at [email protected] for further troubleshooting help.



7 WEEE/RoHS Compliance Statement

EU Directives WEEE and RoHS

To Our Valued Customers:

ibidi GmbH is committed to being a good corporate citizen. As part of that commitment, we

strive to maintain an environmentally conscious manufacturing operation. The European

Union (EU) has enacted two directives, the first on product recycling (Waste Electrical and

Electronic Equipment, WEEE) and the second on limiting the use of certain substances

(Restriction on the use of Hazardous Substances, RoHS).

Two Categories of products covered by the WEEE Directive are currently exempt from the

RoHS Directive – Category 8, medical devices (with the exception of implanted or infected

products) and Category 9, monitoring and control instruments.

All of our products fall into either Category 8 or 9, and are currently exempt from the RoHS

Directive. We will, however, continue to monitor the application of the RoHS Directive in

relation to our products and will comply with any changes, as they apply.

Recycling is offered for our products that fall within the scope of the WEEE Directive. These

specific products, available for sale after August 13, 2005, will be identified with a “wheelie

bin” symbol.

When you see this symbol, do not dispose of this product with Municipal Waste. Special

Collection/Disposal is required.

Please contact us for feedback or further information.

ibidi GmbH

Am Klopferspitz 19

82152 Planegg-Martinsried

Germany

www.ibidi.com

[email protected]

Tel.: +49 89 520 4617 0

Fax: +49 89 520 4617 59



Ibidi Heating System 1, Instruction Manual

© ibidi GmbH 2012, version 2.1, revision 1, 2012-12-03, Dr. Miro Kroutvar, Dr. Zeno Guttenberg I

Appendix

A Menu Navigation

Most of the functions and parameters of the ibidi Temperature Controller can be accessed

with the control buttons on the front panel. The LC display shows the changes. However,

the display is limited to 20 characters per line and four lines. Therefore, some of the fields

are out of the initial view. Nevertheless, they are reachable by using the ’Up / Right’ and

’Down / Left’ arrow buttons. The view shifts accordingly.

By default, there are four channels displayed, but only three of them fit into the viewing

frame. Therefore, to reach the last channel you need to push the ’Up / Right’ arrow button

once more, after you have reached the Channel 3. Then, Channel 4 is displayed and Channel

1 vanishes. By using the ’Down / Left’ arrow button, you will reach Channel 1 again.

A.1 Setting the Temperature

By pressing the control buttons, the cursor on the display will change position. When you

are at the parameter you want to change, you need to select the field with the round ‘Select’

button and change the value using the ‘Up / Right’ and ‘Down / Left’ arrow buttons. To

confirm the change, you need to press the ‘Select’ button again.

Channel names

Measuredte

mperature

Set

temperature

Cursor

Externaltemp

eraturesenso

r

Setup

functions

Run /

Stopbutton

Indicatorfor

additional

view Up / Right

Select / Confirm

Down / Left


II ©ibidi GmbH 2012, version 2.1, revision 1, 2012-12-03, Dr. Miro Kroutvar, Dr. Zeno Guttenberg

A.2 External Temperature Sensor

The ibidi Temperature Controller has plugs for two different temperature sensors:

(a) Type-K plug for a thermo-couple sensor

(b) Plug for a 4-terminal sensing of a Pt100 resistor

You can choose which sensor you want to have displayed and use it as the reference

sensor:

• Thermo-couple: ‘TC-K’

• Pt100 sensor: Pt100

• No selection: ‘NoSel’

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS 42.0 37.0 37.3 Off

E [25.3] Run Setup


E <Pt100>Run Setup


E <TC-K>Run Setup


E <NoSel>Run Setup

A.3 Run / Stop Button

With the ‘Run / Stop’ button, you can set all control channels to standby.

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS [42.0] 37.0 37.3 OffE 25.3 Run Setup

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS 42.0 [37.0] 37.3 OffE 25.3 Run Setup

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS 42.0 37.0 [37.3] OffE 25.3 Run Setup

…

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS 42.0 <37.0> 37.3 OffE 25.3 Run Setup

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS 42.0 <37.1>37.3 OffE 25.3 Run Setup

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS 42.0 <37.2>37.3 OffE 25.3 Run Setup


© ibidi GmbH 2012, version 2.1, revision 1, 2012-12-03, Dr. Miro Kroutvar, Dr. Zeno Guttenberg III

A.4 Setup Button

Using the ‘Setup’ button, you can reach further functions, such as the channel output

‘Mode’, the ‘Alarm’ function, the controller ‘Preferences’, the ‘Calibration’ function, and you

can view the ’Info’ (information) about the controller.

A.4.1 Mode Function

In this field, you can switch the channels individually ‘On’ and ‘Off’. For that, you have to

select the corresponding channel number and change the ‘Mode’ from ‘On’ to ‘Off’.

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 Off

S 42.0 37.0 37.3 Off

E 25.3 [Run] Setup

Ch1 Ch2 Ch3 Ch4

I 42.1 36.9 37.3 Off

S 42.0 37.0 37.3 OffE 25.3 <Run> Setup

Ch1 Ch2 Ch3 Ch4

I 42.1 36.9 37.3 Off

S 42.0 37.0 37.3 OffE 25.3 <Stop> Setup

Setup <Mode>

Channel = 1Mode = On

Return

Setup <Alarms>

Channel = 1High = +40.0 °CLow = +25.5 °C

Return

Setup <Preferences>

Backlight = 65Contrast = 25

Return

Setup <Calibration>Channel 1 Corr +0.0

Calibrate nowMeas +42.1 Ref +41.5

Return

Setup <Info>TempCoSN: xxxxFirmware 1.01

Return

Setup <Return>

…

…

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS 42.0 37.0 37.3 OffE 25.3 Run [Setup]

Ch1 Ch2 Ch3 Ch4I 42.1 36.9 37.3 OffS 42.0 37.0 37.3 OffE 25.3 Run [Setup]

Setup <Mode>


Return

Setup <Alarms>


Return

Setup <Mode>


Return

Setup <Alarms>


Return

Setup <Alarms>


Return

Setup <Mode>


Return

Setup <Alarms>


Return

Setup <Mode>


Return

Setup <Alarms>


Return

Setup <Info>TempCo

SN: ibiTC-xxxFirmware 0.35E5

Return

Setup <Return>


IV ©ibidi GmbH 2012, version 2.1, revision 1, 2012-12-03, Dr. Miro Kroutvar, Dr. Zeno Guttenberg

Setup Mode

Channel = [1]Mode = On

Return

Setup Mode

Channel = 1Mode = [On]

Return

Setup Mode


[Return]

Setup ModeChannel = <1>

Mode = On

Return

Setup ModeChannel = 1

Mode = <On>

Return

Setup ModeChannel = 1

Mode = <Off>

Return

Setup <Mode>Channel = 1

Mode = OnReturn

A.4.2 Alarm Function

If you activate the ‘Alarm’ function, you will get a visual (�’High’ or ‘Low’ message on the

LCD) and optionally acoustical signal (� Beeper, please contact ibidi). The alarm goes off

when the measured temperature exceeds the upper or lower thresholds of the alarm limits.

The limits are individually programmable. You can set the thresholds as absolute or relative

temperatures using the TempControl software.

Setup AlarmsChannel = [1]

High = +40.0 °CLow = +25.5 °C

Return

Setup AlarmsChannel = 1

High = [+40.0] °CLow = +25.5 °C

Return


High = +40.0 °CLow = [+25.5] °C

Return


High = +40.0 °CLow = +25.5 °C

[Return]

Setup AlarmsChannel = <1>High = +40.0 °CLow = +25.5 °C

Return

Setup AlarmsChannel = 1High = <+40.0>°CLow = +25.5 °C

Return

Setup AlarmsChannel = 1High = <+40.1>°CLow = +25.5 °C

Return

Setup AlarmsChannel = 1High = +40.0 °CLow = <+25.5>°C

Return

Setup <Alarms>Channel = 1High = +40.0 °CLow = +25.5 °C

Return


© ibidi GmbH 2012, version 2.1, revision 1, 2012-12-03, Dr. Miro Kroutvar, Dr. Zeno Guttenberg V

A.4.3 Preferences

Setup <Preferences>

Backlight = 65

Contrast = 25Return

Setup PreferencesBacklight = [65]

Contrast = 25

Return

Setup PreferencesBacklight = 65

Contrast = [25]

Return

Setup PreferencesBacklight = 65

Contrast = 25

[Return]

Setup Preferences

Backlight = 65

Contrast = <25>Return

Setup Preferences

Backlight = 65

Contrast = <26> Return

Setup PreferencesBacklight = <65>

Contrast = 25

Return

Setup PreferencesBacklight = <66>

Contrast = 25

Return

The display settings of the controller can be changed under ‘Preferences’. You can modify

the brightness of the display (‘Backlight’) and the contrast.

A.4.4 Calibration

The ibidi Temperature Controller has a built-in function for calibrating the controlled channels.

To do this, there are two plugs for external temperature sensors on the rear panel of the

controller. In this menu, you will find the channel number, the currently measured

temperature ‘Meas’, the sensed temperature ‘Ref’ of the external sensor, and the currently

used temperature offset ‘Corr’. You can change the reference sensor, as described in

Section A.2.


VI ©ibidi GmbH 2012, version 2.1, revision 1, 2012-12-03, Dr. Miro Kroutvar, Dr. Zeno Guttenberg

Setup <Calibration>Channel 1 Corr +0.0Calibrate now

Meas +42.1 Ref +41.5Return

Setup Calibration

Channel [1]Corr +0.0Calibrate nowMeas +42.1 Ref +41.5

Return

Setup CalibrationChannel 1 Corr +0.0

Calibrate [now]Meas +42.1 Ref +41.5

Return

Setup Calibration

Channel 1 Corr +0.0Calibrate nowMeas +42.1 Ref +41.5

[Return]

Setup CalibrationChannel <1> Corr +0.0Calibrate now


Setup CalibrationChannel<2>Corr +0.0Calibrate now


Setup CalibrationChannel 1 Corr -0.6Calibrate now


automatic

Once you have activated the calibration ‘Calibrate Now’, the controller will calculate an offset

from the temperature difference between the control channel and the external sensor. This

offset will replace the former value of ‘Corr’.

A.4.5 Info

Setup <Info>TempCoSN: ibiTC-xxxFirmware 0.31E5

Return

In this menu, you can find the serial number of the device, as well as the hardware and

firmware versions.

Documents

Instruction Manual for the ibidi Heating System 1 ... · Instruction Manual for the ibidi Heating System 1, Universal Fit Version 2.1 ... variety of optical microscope setup requirements