Okolab H301-T-UNIT-BL-PLUS User manual

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H301-T-UNIT-BL-PLUS
Electric Top Stage Incubation System
Manual
Vers. 01.17

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Index
INDEX
1PREFACE ............................................................................................................................................................ 5
2SYMBOLS DESCRIPTION .................................................................................................................................... 6
2.1 Symbols in this manual and on the product ...............................................................................6
2.2 Symbols on the product label ................................................................................................6
3SAFETY NOTES .................................................................................................................................................. 7
4SYSTEM OVERVIEW ........................................................................................................................................... 9
4.1 Equipment supplied............................................................................................................9
4.2 Equipment not supplied.......................................................................................................9
4.2.1 Equipment Required........................................................................................................9
4.2.2 Optional ......................................................................................................................9
4.3 Temperature Controller connection ports ............................................................................... 10
5H301-T-UNIT-BL-PLUS. INSTALLATION AND USER GUIDE ......................................................................... 10
5.1 H301-T-UNIT-BL-PLUS. Installation guide ................................................................................ 10
5.2 How to connect H301-T-UNIT-BL-PLUS to a BOLD LINE Digital Gas Controller .................................... 13
6USER INTERFACE............................................................................................................................................. 15
6.1 Home page .................................................................................................................... 15
6.1.1 Colors meaning ............................................................................................................ 15
6.1.2 Changing Set Points....................................................................................................... 16
6.2 Settings ........................................................................................................................ 16
6.2.1 Temperature............................................................................................................... 16
6.2.1.1 Chamber/Adapter............................................................................................................. 16
6.2.1.2 Control Mode ..................................................................................................................... 17
6.2.1.3 Objective Heater .............................................................................................................. 21
6.2.1.4 Thermal Cycles.................................................................................................................. 24
6.2.1.5 Open Incubator ................................................................................................................. 26
6.2.1.6 Ramp Rate.......................................................................................................................... 27
6.2.1.7 External Sensors................................................................................................................ 27
6.2.1.8 Status .................................................................................................................................. 28
6.2.1.9 Digital Input....................................................................................................................... 28
6.2.2 Touch Screen .............................................................................................................. 29
6.2.2.1 Options ............................................................................................................................... 29
6.2.2.2 Brightness........................................................................................................................... 30
6.2.2.3 Calibration ......................................................................................................................... 30
6.2.2.4 Date & Time....................................................................................................................... 30

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6.2.2.5 Alarms................................................................................................................................. 31
6.2.2.6 Visual Effects..................................................................................................................... 31
6.3 Overview Page ................................................................................................................ 32
6.4 Glance Mode .................................................................................................................. 32
7CALIBRATIONS................................................................................................................................................. 34
7.1 Chamber Self-Calibration ................................................................................................... 34
7.2 Objective Heater Manual Calibration ..................................................................................... 35
7.3 Objective Heater Self-Calibration......................................................................................... 37
8MAINTENANCE ................................................................................................................................................. 39
9SUPPORT.......................................................................................................................................................... 40
9.1 QR Codes for Support........................................................................................................ 40
9.2 Support Contacts ............................................................................................................. 40
9.3 Web Conference For Assistance And Training ........................................................................... 40
10 TROUBLESHOOTING.................................................................................................................................. 42
11 TECHNICAL SPECIFICATIONS ................................................................................................................... 43
12 FIGURE LIST ............................................................................................................................................... 44
13 REVISION TABLE ........................................................................................................................................ 45

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1PREFACE
H301-T-UNIT-BL-PLUS is a temperature controller that can operate any Okolab Electrically Heated Stage
Top Incubator to maintain the required thermal conditions for live cell imaging right on the microscope
stage. H301-T-UNIT-BL-PLUS is compatible with Okolab Objective Heaters and Digital Gas Controllers to
maintain the sample in physiological condition with the highest accuracy, thus allowing long term time lapse
imaging.
H301-T-UNIT-BL-PLUS controls the temperature of the incubator base and lid and, when connected, of the
Objective Heater. To improve accuracy on sample temperature, H301-T-UNIT-BL-PLUS is equipped with a
flexible fine gauge thermocouple that can be used to provide feedback from the temperature of a reference
well located in close proximity to the experimental sample. H301-T-UNIT-BL-PLUS is equipped with a room
temperature sensor to monitor laboratory temperature.
H301-T-UNIT-BL-PLUS can be operated via the convenient touch screen interface OKO-TOUCH (not
included), via OKOLAB DataLog Software or via any Third Party Software.

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2SYMBOLS DESCRIPTION
The following symbols identify important information to note:
CAUTION or WARNING: This symbol warns you about the risk of electrical shock.
CAUTION or WARNING or IMPORTANT: This symbol warns you of circumstances or practices
that can affect the instrument’s functionality. Please refer to accompanying documents.
Tip -Supplies you with helpful suggestions.
Note -Supplies you with important information to successfully setup and use the instrument.
CE MARKING: This symbol indicates a product’s compliance with EU legislation.
PRODUCT DISPOSAL: This symbol indicates that this product must not be disposed as urban
solid waste.
This symbol indicates the product production date
This symbol indicates the manufacturer data
This symbol indicates the protection degree against ingress of solids or liquids inside the
product.
This symbol warns you to read the user manual before starting the device

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3SAFETY NOTES
Before operating the equipment please read carefully the instructions and the safety notes. If you have any
questions, please contact Okolab.
The equipment must only be used as intended and as described in this Manual.
Equipment should only be operated by technically qualified personnel.
Do not start up the equipment if some of its parts are damaged.
This instrument is not intended for use in locations where flammable or explosive gases are
present.
Handle the equipment with care.
Equipment and its internal parts can be damaged by dropping and by shock.
Do not use a volatile solvent such as paint thinner to clean the instrument. Deformation or
discoloration will occur. Use a soft, dry cloth to remove stains from the instrument. Not
following these instructions can result in damage or breakdown of the device and its
accessories.
The products labels can be found on the bottom panel of the Main Unit.
Do not exceed voltage indicated in this manual and on the product label.
Avoid excessive induction noise, static electricity, magnetic fields.
Do not expose this instrument to rain or moisture.
Prevent throttling and kinking of tubing.
Check the cables are well fixed to their own connectors so they cannot slip off
This device is not designed for use under medical conditions.
Power cord of unit should be unplugged from electrical outlet when left unused for long period
of time.
VENTILATION, unit should be situated so that its' location or position does not interfere with
proper ventilation.
Unit should be situated away from heat sources such as radiators, heat registers,
stoves, or other appliances or processes that produce heat
Do not start up the equipment if the supply cable is damaged.
Connect the equipment only to grounded mains power socket.
Do not disconnect cables while in operation.
Do not open the unit. Do not remove cover or back.

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Prevent metal fragments or lead wire scraps from falling inside instrument case to avoid
electric shock, fire or malfunction.
No user serviceable parts inside.
Before starting, assemble the equipment while unplugged from an outlet.
Unit should never be used where it can fall or be pushed into water.
International caution symbol marks this device. IMPORTANT: read the “Safety Notes”
before installing, using and commissioning this device, as the notes contain important
information relating to safety and EMC. Not following these instructions can result in
damage or breakdown of the device and its accessories.
We reserve the right to make technical variations.
IN NO EVENT SHALL OKOLAB S.R.L. BE LIABLE FOR ANY DIRECT, INCIDENTAL OR CONSEQUENTIAL DAMAGES
OF ANY NATURE, LOSSES OR EXPENSES RESULTING FROM ANY DEFECTIVE PRODUCT OR THE USE OF ANY
PRODUCT.

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4SYSTEM OVERVIEW
H301-T-UNIT-BL-PLUS includes:
1. T-Unit
2. Fine Gauge Thermocouple. To measure specimen temperature
3. NTC Thermistor. To measure ambient temperature
4. #1 24V-DC Power Adapter
4.2.1 Equipment Required
•Okolab incubating chamber with heated lid and base, chamber riser with one or more plate
adapters
4.2.2 Optional
•OKO-TOUCH, including:
1. Touch Screen Interface
2. #1 5V-DC Power adapter. To connect the Touch Screen.
3. #1 RS232 Serial cable. To connect the Touch Screen to the T-Unit
•Objective heater. Required ONLY when using immersion or dipping objectives.

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Figure 1: Control Unit Rear Panel Overview
1. OKO-TOUCH RS232 serial port.
2. Power Input.
3. Chamber base connector.
4. Lid chamber connector.
5. Objective Heater connector (only if you have ordered also an OBJ-COLLAR-####)
6. USB port.
7. General Purpose Input Output (GPIO) port.
8. NTC Thermistor port.
9. Gauge Thermocouple port.
5H301-T-UNIT-BL-PLUS. INSTALLATION AND USER GUIDE
The following paragraphs illustrate how to install and use the Temperature Control Unit.
The following instructions will guide you through a quick installation. For safe operation of the
unit it is absolutely necessary to read carefully all the instructions and safety notes.

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Figure 2. H301-T-BL-PLUS. Standard Connection
1. Connect the base of the incubation chamber to the port labeled ‘Base’ on the rear panel of the
Temperature Control Unit (see Figure 2).
2. Connect the lid of the incubation chamber to the port labeled ‘Lid’ on the rear panel of the
Temperature Control Unit (see Figure 2).
3. Plug the NTC Connector on the port labeled ‘Ambient Temperature’ located on the rear panel of the
Temperature Control Unit (see Figure 2). This temperature sensor must be placed in air close to the
illumination pillar in proximity to the microscope stage. This thermistor is monitoring the room
temperature.
Note ►Stage Top Chambers for most brands/models are available. See www.oko-lab.com for details.
4. The Fine Gauge Thermocouple (green) plugs into the green port labeled ‘Reference Temperature’
located on the rear panel of the Temperature Control Unit (see Figure 2). When operating the device
in ‘Sample Feedback Mode’ place this thermocouple in a Reference Well filled with water. Make sure
that the end of the Fine Gauge Thermocouple is fully immersed.
Tip ►The Fine Gauge Thermocouple may be placed into a Reference Well if running the “Sample Feedback
Mode” (see paragraph 6.2.1.2). The Fine Gauge Thermocouple may also be used to calibrate the Stage
Top Incubation Chamber and/or the Objective Heater (OBJ-COLLAR ####), when present. The Fine
Gauge Thermocouple may be left in air if the system is operated in ‘Chamber feedback mode’ (see
paragraph 6.2.1.2)

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5. When using oil immersion objectives an Objective Heater should also be purchased. If your system is
also equipped with the Objective Heater, (OBJ-COLLAR ####, optional) its objective collar should be
installed before connecting the heater to rear panel of the Temperature Control Unit. Follow the
steps shown in Figure 3. Once you have assembled it, plug it into the port labeled ‘Obj. Heater’
located on the rear panel of the Temperature Control Unit (see Figure 4).
Figure 3.Objective Heater (OBJ-COLLAR ####) Assembly
Figure 4. OBJ-COLLAR Connection (Optional)
6. Connect the touch screen interface OKO-TOUCH to the Temperature Control Unit using the serial
cable provided (see Figure 5).
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Objective Band Heater
Velcro Tape

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Figure 5. OKO-TOUCH Connection
7. Connect the power adapter to OKO-TOUCH. Connect the additional power cord to the Temperature
Control Unit.
8. Turn the OKO-TOUCH on (rocking switch is located on the back panel of OKO-TOUCH). A small green
LED light will blink on the front of the Temperature Control Unit. This means that the system is
initializing.
Once the system has been initialized the LED on the Temperature Control Unit will no longer blink and will
be steadily on. The OKO-TOUCH interface will display its homepage and the indicator light on the
temperature tab will be initially yellow. When the temperature set-point is within the set tolerance range,
the indicator light will turn green.
Tip ►Wait until system has reached the steady state before loading your samples.
The H301-T-UNIT-BL-PLUS can be used as a stand-alone device or in combination with any one of the Bold
Line Gas Controllers. Figure 6 shows how to assemble the H301-T UNIT-BL-PLUS, for example, with a CO2-
O2 Unit BL [0-10; 1-18].

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a. CO2-O2 UNIT-BL [0-10; 1-18] and
H301-T-UNIT-BL-PLUS. Units rear panels.
b. CO2-O2 UNIT-BL [0-10; 1-18] and
H301-T-UNIT-BL-PLUS. 3D view.
Figure 6. CO2-O2 UNIT-BL [0-10; 1-18] and H301-T-UNIT-BL-PLUS
The T-Unit must stack on top of the Gas Controller by lining up the bus ports located on the top and bottom
surface of each. When the units are properly connected the Temperature, CO2 and O2 parameters will
appear on the OKO-TOUCH Touch Screen interface Home page. If not all the parameters appear on OKO-
TOUCH Home Page then the Control Units are not properly connected. Please check that the bus ports are
properly aligned with each other.
If you have a Gas Controller and a Temperature Control Unit, it is essential to read both this
manual and the Gas Controller Manual to familiarize yourself with the functions and the
operation of the devices before use.

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6USER INTERFACE
Figure 7. Homepage of the Control Unit Touch Screen Display
On the right side of the screen you will see the graph of Temperature value over time. The X axis displays
the scale of the duration of time you selected (in this example 1 hour). The Y axis, shows the temperature
scale.
Tip ►The Touch Screen is pre-set at the following temperature: 37°C. Once turning the Touch Screen on it
will start operating to reach this set-point value.
6.1.1 Colors meaning
The GREEN indicator means that set-point value has been reached (within the tolerance
you’ve set) and that the system is working properly.
The YELLOW indicator means that the system is working towards reaching set points. NO
actions on your part are required. The Yellow light will also appear every time you change
the Temperature set-point.
The ORANGE indicator means that the current gas concentration is not correct and its value
is out of the set tolerance (see section “ALARMS” in paragraph 6.2.2.5). While giving time to
the system to recover from this out-of-range value, please double check if there may be
reasons not depending on the unit for which it is not properly working. Verify if all cables are
correctly connected.
Maximum T °C reached within the time-period
Temperature set-point
Touch here to access system
options and settings
Touch here to go to
the OVERVIEW page
Current
Temperature value
Chart history lenght
set at 1 h
Temperature chart
Minimum T °C reached within the time-period
Touch here to know
generic info about Control
Unit and running time

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The RED indicator means that there could be a problem with the unit. If after 20-30 minutes
the Red light does not turn to Green, turn the system off, wait 5 minutes and turn it back on.
If the problem persists contact Okolab at http://www.oko-lab.com
6.1.2 Changing Set Points
To change a set point, simply touch the corresponding tab. A new page will open where you can press +/–
to adjust the temperature to the desired value from 3°C above ambient to 60°C. Press “Set” to save the
new set point or “Cancel” to undo.
Figure 8. How to change the Temperature set point
6.2.1 Temperature
Temperature Settings are adjusted by following the two steps below (look at the touching finger)
Figure 9. Temperature settings
6.2.1.1 Chamber/Adapter
Figure 10. Here you have to set the model of incubating chamber (it depends on your XY stage/piezo) that
you have purchased and the type of sample adapter being inserted into it. Depending on these selections,
the thermal controller will automatically change settings. To switch configuration press on the Chamber or
on the Adapter icon (see Figure 10), then scroll among different chambers/adapters using the arrows on the
left and right sides of the screen. If you choose to change the chamber, when the chamber you are about
to use appears on the screen press “Next”. Repeat this sequence to find and select the sample adapter (see
Figure 11) and press “Ok”(see Figure 12). The unit will save these settings.
1
2

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Figure 10. Chamber & Adapter change
Figure 11 Chamber & Adapter settings
Figure 12. Chamber and adapters saving
Tip ►The system is shipped from the factory already set according to the incubating chamber model and
sample adapter you have ordered. Keep in mind that thermal control settings change when changing the
adapter in use. Hence, every time you change adapter you must input the correct one using the procedure
above. In case you purchase additional incubating chamber models make sure to input the correct
chamber/sample insert in the Setting prior to your experiment.
6.2.1.2 Control Mode
The Temperature Control can run in two different modes, please choose which one you want to use, Sample
or Chamber Feedback Mode.
Figure 13 shows how to select Chamber Feedback Mode.

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Figure 13. Chamber Mode Setting
In this configuration the temperatures of both the lid and the base of the incubating chamber are strictly
controlled. A careful calibration performed in our laboratories between these temperatures and that of the
sample, guarantees that sample temperature is maintained at the desired set point value. The advantage
of this solution is that you don’t have to manually place the Fine Gauge Thermocouple into a Reference
Well.
Tip ►Advantages of this Control Mode are: fast experiment start up and no specific action required in multi
user applications. The disadvantage is that it needs calibration in case of room temperature variations
beyond +/- 1 degree Celsius.
When selecting Chamber Feedback Mode the following Screen will appear:
Figure 14. Chamber calibration start
It states that the system has been calibrated at the factory at a Room Temperature of 23°C. In case your
Room Temperature, measured by the included thermistor, is different from 23°C (i.e. 20.3°C) you have 3
options:
1. Use Current Calibration: this means recalling the last saved calibration settings. Once you calibrate
the system (see point 3 below) these settings will be automatically stored. The system saves only
the last calibration performed. Please note that calibration depends on i) set-point temperature; ii)
room temperature; iii) type of sample adapter in use. Hence using ‘Custom Calibration’ is NOT
recommended in any of the following as it will yield inaccurate results:
A. If the recalled calibration was performed with a different sample adapter than what you intend
to use
B. If the room temperature on the Custom Calibration is significantly different than the current one
(i.e. more than 1°C)

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C. If the Custom Calibration was performed with a different set point temperature than what you
plan on using (e.g Custom Calibration had set point of 37C but new experiment requires a set point
temperature of 39C).
2. Use Factory settings optimized at 23°C Room Temperature. This is acceptable if your room
temperature is within 1°C from 23°C. However, for room temperature more than 1°C from 23°C this
may be not acceptable depending on the sensitivity of your sample.
3. Choose to Calibrate the system in your lab at your Room Temperature, for maximum of accuracy
when using “Chamber Feedback Mode”. To do so follow the instructions on paragraph 7 (Chamber
Self-Calibration) and then press the “Calibrate” button; the automatic calibration procedure will
start.
Tip ►Note please it may take 1-2 hours to complete the calibration.
Figure 15. Chamber calibration progress
Just wait until the following dialog box appears:
Figure 16. Chamber Calibration result
Figure 17 shows how to select the Sample Feedback Mode.

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Figure 17. Sample Mode setting
In this configuration, every time you run an experiment, you must place the Fine Gauge Thermocouple (thin
green wire) into a Petri dish or well adjacent to your sample (from here on this will be referred to as
Reference Well). Fill the Reference Well with distilled water to a level allowing full immersion of the fine
gauge thermocouple (see Figure 49). When possible close the petri with its own plastic lid (or with SENSOR
LID, see Tip in paragraph 7) and place it in the appropriate holder. Using a Reference Well in close proximity
to your Experimental Well provides active Sample Feedback Temperature Control. This ensures that your
sample will be within 0.1°C from the temperature set point regardless the room temperature variations.
Tip ►Advantages of Sample Feedback Control Mode are: 1) direct monitoring and active control of sample
temperature and 2) independence from room temperature variations. The disadvantage is that it requires
manual operation for correct placement of the fine gauge thermocouple in a Reference Well.
The Control Mode page also includes an Open Incubator button (see Figure 18). Press Open Incubator before
opening the lid of the Incubation Chamber (e.g. in order to add culture medium). This places the thermal
controller in stand-by and avoids overheating the sample.
Note ►: this feature is available only when operating in Sample Feedback Mode Control.
Figure 18. Open Incubator
After closing the incubation chamber wait a couple of minutes and press “Incubator Closed”. Now you can
continue your experiment.