Okolab Bold Line H101-CRYO-BL User manual

1
H101-CRYO-BL
Manual

2
This page was left blank

3
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 required............................................................................................................9
4.3 Optional equipment............................................................................................................9
4.4 T-Unit connection ports..................................................................................................... 10
5H101-CRYO-BL. INSTALLATION AND USER GUIDE...................................................................................... 10
5.1 How to connect the T-Unit to OKOLAB BOLD LINE Gas Controller................................................... 13
6USER INTERFACE............................................................................................................................................. 14
6.1 Home page .................................................................................................................... 14
6.1.1 Colors meaning ............................................................................................................ 14
6.1.2 Changing Set Points....................................................................................................... 15
6.2 Settings ........................................................................................................................ 15
6.2.1 Temperature............................................................................................................... 15
6.2.1.1 Control Mode ..................................................................................................................... 15
6.2.1.2 Objective Heater (used only in exceptional cases) .................................................... 18
6.2.1.3 Thermal Cycles.................................................................................................................. 19
6.2.1.4 Status.................................................................................................................................. 21
6.2.1.5 External Sensors................................................................................................................ 22
6.2.1.6 Digital Input....................................................................................................................... 22
6.2.2 Touch Screen .............................................................................................................. 23
6.2.2.1 Options ............................................................................................................................... 23
6.2.2.2 Brightness........................................................................................................................... 23
6.2.2.3 Visual Effects..................................................................................................................... 24
6.2.2.4 Date & Time....................................................................................................................... 24
6.2.2.5 Alarms................................................................................................................................. 25
6.2.2.6 Logging ............................................................................................................................... 25
6.3 Overview Page ................................................................................................................ 25
6.4 Glance Mode .................................................................................................................. 26
7WATER BATH-CRYO APPENDIX ..................................................................................................................... 27
7.1 Menu function................................................................................................................. 27
7.2 Thermostatic Water Bath parameters .................................................................................... 27

4
7.3 Display scheme ............................................................................................................... 27
7.4 Adjustment of the pump outflows ........................................................................................ 29
7.5Pump output .................................................................................................................. 29
7.6 Standard Cooling value...................................................................................................... 30
8MAINTENANCE OF THE T-UNIT ..................................................................................................................... 31
9WATER BATH MAINTENANCE......................................................................................................................... 31
10 SUPPORT .................................................................................................................................................... 33
10.1 QR Codes for Support........................................................................................................ 33
10.2 Support Contacts ............................................................................................................. 33
10.3 Web Conference For Assistance And Training ........................................................................... 33
11 TROUBLESHOOTING.................................................................................................................................. 34
12 TECHNICAL SPECIFICATIONS ................................................................................................................... 35
13 FIGURE LIST ............................................................................................................................................... 36

5
1PREFACE
The H101-CRYO-BL, is designed to maintain all the thermal conditions required around your microscope,
thus enabling to carry out prolonged observations on biological specimens. The H101-CRYO-BL controls the
temperature in the range from 10/15°C below Room temperature (depending on the chamber model) to
60°C. The H101-CRYO-BL is composed of the T-Unit (Temperature Control Unit), the Water Bath CRYO
(heating and cooling), a fine gauge thermocouple to monitor the temperature of a blank sample or the
Reference Well located in close proximity to the experimental sample, a thermistor to monitor the Room
temperature. The H101-CRYO-BL needs to be connected with the Okolab incubating chamber with one or
more plate adapters
The H101-CRYO-BL can be operated via the touch screen interface OKO-TOUCH (not included), via OKOLAB
DATA-LOG Software (not included) or via any Third Party Software.
The H101-CRYO-BL is compatible with any Okolab Bold Line Gas Controller. In this case, a single OKO-TOUCH
operates both, the Bold Line Gas Controller and T-Unit.
The H101-CRYO-BL is compatible with OKO-MOC (not included).

6
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

7
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 T-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.

8
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.

9
4SYSTEM OVERVIEW
H101-CRYO-BL includes:
1. T-Unit
2. Water Bath CRYO (heating and cooling)
3. 3 PIN CABLE –RS232. To connect the Water Bath to the T-Unit
4. Fine Gauge Thermocouple. To measure specimen temperature
5. NTC Thermistor. To measure ambient temperature
6. TUBE-T - Blue marked tubing composed of transparent silicon tube 2mm ID (to be connected to the
colour-related connector of the incubating chamber) connected with a 10mm ID black butyl tube
isolated (to be connected to the colour-related connector of the water-bath)
7. TUBE-S -Red marked tubing composed of transparent silicon tube 2mm ID (to be connected to the
colour-related connector of the incubating chamber) connected with a 10mm ID black butyl tube
isolated (to be connected to the colour-related connector of the water-bath)
8. #1 24V-DC Power Adapter
•Okolab incubating chamber with lid and base, chamber riser with one or more plate adapters
•OKO-TOUCH, includes:
-RS232 Serial cable. To connect the Touch Screen to the T-Unit
•OKO-MOC

10
2 3 4 5 6 7 8
1
Figure 1: T-Unit Rear Panel Overview
1. OKO-TOUCH RS232 serial port.
2. Power Input.
3. Objective Heater (optional) connector.
4. Water Bath CRYO connector.
5. USB port.
6. General Purpose Input Output (GPIO) port.
7. NTC Thermistor port.
8. Gauge Thermocouple port.
5H101-CRYO-BL. INSTALLATION AND USER GUIDE
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.

11
Figure 2. H101-CRYO-BL Standard Connection
1. Connect the Water Bath CRYO to the port labeled ‘Water Bath’ on the rear panel of the T-Unit (see
Figure 2).
2. Plug the NTC Connector on the port labeled ‘Ambient Temperature’ located on the rear panel of the
T-Unit (see Figure 2). The NTC thermistor can be also used to monitor the room temperature when
working in Sample Feedback Mode.
3. The Fine Gauge Thermocouple (green) plugs into the green port labeled ‘Reference Temperature’
located on the rear panel of the T-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 6.2.1.1). The Fine Gauge Thermocouple may be left in air if the system is
operated in ‘Chamber feedback mode’ (see paragraph 6.2.1.1).
4. Connect the touch screen interface OKO-TOUCH to the T-Unit using the serial cable provided (see
Figure 3).

12
Figure 3. OKO-TOUCH Connection
5. Connect the TUBE-S and TUBE-T between Okolab Chamber Incubator and the Water Bath CRYO (see
Figure 4).
Figure 4. Tubing connection between Okolab Chamber and the Water Bath CRYO
6. Connect the 24V-DC Power adapter to the T-Unit.
7. To switch the T-Unit on use the power button on OKO-TOUCH. A small green LED light will blink on
the front of the T-Unit. This means that the system is initializing.
Once the system has been initialized the LED on the T-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

13
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 T-Unit can be used as a stand-alone device or in combination with any one of the Bold Line Gas
Controllers. Figure 5 shows how to assemble the T-Unit, for example, with a CO2-O2 Unit BL [0-10; 1-18].
a
b
Figure 5. CO2-O2 UNIT-BL [0-10; 1-18] and T-UNIT connection (a-b)
The Gas Controller must stack on top of the T-Unit 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 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.

14
6USER INTERFACE
Figure 6. OKO-TOUCH Home page
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 OKO-TOUCHT 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

15
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 7. 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 8. Temperature settings
6.2.1.1 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 9 shows how to select Chamber Feedback Mode.
1
2

16
Figure 9. Chamber Mode Setting
In this configuration the water bath temperature is controlled. For maximum of accuracy, once selected
the Chamber feedback mode, the operator should calibrate the difference of temperature between the
water bath temperature and the specimen temperature (by sticking the fine gauge thermocouple into a
reference well inside the Chamber Incubator), so to find for instance that to keep the specimen temperature
at 37°C, the water bath temperature has to be at 40°C. This means that when regulating the set-point,
given that in this mode you control the Chamber’s temperature, this has to be set at 40°C to keep the
specimen at 37°C. This control mode is well suited for multi-user tasks where users don’t want to take care
to correctly position the fine gauge thermocouple into a reference well inside the chamber incubator;
however it is based on a manual calibration between the water bath and the specimen, therefore cannot
be considered an active thermal control on the sample
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 between water bath and the specimen. The
temperature difference between the water bath and the sample sensor depends on many factors like: the
ambient temperature, the enclosure design, the sensors position, but if these ones don’t change, the
temperature difference doesn’t change.
Figure 10 shows how to select Sample Feedback Mode.
Figure 10. 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 11). When possible close the petri with its own plastic lid (or with SENSOR
LID, see Tip below) and place it in the appropriate holder. Using a Reference Well in close proximity to your

17
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 from room temperature variations.
Figure 11. Free Sensor attached on the internal base of the Petri Dish
Tip ►It is recommended using an Okolab SENSOR LID (available for purchase) to secure the Fine Gauge
Thermocouple in place in the Reference Well during calibration or operation in Sample Feedback Mode (see
Figure 12).
Sensor lids are available for the following: 35 mm, 60 mm Petri-dish, chamber slide and chambered cover
glass. Sensor Lids are ordered separately using the codes in following table.
SENSOR LID CODES
SENSOR LID-35
Temperature sensor lid. To be used in specimen feedback in a 35 mm Petri Dish
SENSOR LID-60
Temperature sensor lid. To be used in specimen feedback in a 60 mm Petri Dish
SENSOR LID-GS
Temperature sensor lid. To be used in specimen feedback in chamber slides and
chambered cover glass
Table 1. Sensors lid codes
Figure 12. SENSOR LID-35. For 35 mm Petri-dish
Tip ►Advantages of Sample Feedback Control Mode are: direct monitoring and active control of sample
temperature and 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.
Tip ►During the normal operation it could be necessary to open the incubator. The picture below shows
the procedure to follow when opening incubator:
•Press on Open Incubator, before removing the lid. Now you can open the incubator;
SENSOR LID
Fine Gauge
Thermocouple

18
•Once you have done close the chamber and wait for 2 minutes in order to let the environment achieve
steady state; press on Incubator Closed and the temperature controller will resume operating
normally.
Figure 13. Open incubator
Opening the incubator can lead to controller instability since both the specimen and the
inside of the chamber are exposed to the external environment. In order to avoid the
problem, H101-CRYO-BL has to stay in a holding state until the incubator is closed and its
environment is in a steady state. This can be achieved using the Open Incubator procedure.
6.2.1.2 Objective Heater (optional)
Figure 14 shows how to enable/disable the Objective Heater (OBJ COLLAR ####). The Objective Heater is
disabled by default. Read this paragraph only if your system is equipped with Objective Heater (OBJ COLLAR
####).
Figure 14. Objective Heater enabling
Tip ►Before enabling it, make sure it is already installed around your objective, and that it has been
correctly connected to the T-Unit.
When enabling and saving the Objective Heater the following Screenshot will appear:
1
2

19
Figure 15. Objective Heater Calibration Page
The temperature of the Objective Heater is factory calibrated at room temperature of 23°C, with a 3
degrees offset relative to the sample set-point temperature of 37°C. This offset value is appropriate for a
set point of 37°C. If your set point is different than 37°C the system will automatically apply the correct
offset. See Figure 15 above. This means that if the temperature is set at 37°C the Objective Heater is kept
at 40°C in order to maintain the sample at the desired temperature while imaging with an oil immersion
objective. To accept this offset you can press “USE FACTORY” and return to the Homepage. Alternatively
the following options are available:
8. MANUAL: Selecting “MANUAL” allows searching for the best offset value manually, step by step. See
Figure 16.
Figure 16. Objective Heater Manual Offset
6.2.1.3 Thermal Cycles
This function allows the incubator to switch between two different temperatures (both temperatures must
be at least 3°C above Room Temperature).

20
Figure 17. Thermal Cycles Subpage
You can decide whether setting a new thermal cycle (“New”) or recalling a previously saved one (“Load”).
To set a new thermal cycle, first choose the two temperatures between which you want to switch (SP1: Set
Point 1 and SP2: Set Point 2). Next, set the duration in hours and minutes you want the system to dwell on
each temperature. Lastly set the number of thermal cycles (“Number”) you want the system to perform
(see Figure 18)
Figure 18. Thermal cycles settings
Tip ►The system switches from a higher to a lower temperature by dissipation; hence it requires some
time (the amount of time required depends on several factors such as control mode, room temperature and
set-points). Please keep this in mind when setting the “time” between the two set points.
Once you have adjusted the settings, a “Cycle Summary” page will be displayed. Press “Save As” if you want
to save the Thermal Cycle program to be recalled for future experiments. You can save up to 4 different
thermal cycle programs. Saving a 5th one will overwrite the first saved one.
1
3
2
1
2
3