Okolab UNO-COMBINED-CONTROLLER User manual

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UNO-COMBINED-CONTROLLER
CO2Microscope Electric Top Stage Incubation System
Manual
Vers. 03.16

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Index
Index
1REVISION TABLE ............................................................................................................................................ 5
2PREFACE.......................................................................................................................................................... 6
3SYMBOLS DESCRIPTIONS............................................................................................................................ 7
3.1.1 Symbols in this manual and on the product:............................................................................7
3.1.2 Symbols on the product label: ............................................................................................7
4SAFETY NOTES............................................................................................................................................... 7
5AC SECTION WIRING DIAGRAM .................................................................................................................. 9
6UNO-COMBINED-CONTROLLER. DESCRIPTION. ................................................................................... 11
7EQUIPMENT AND CONNECTIONS ............................................................................................................. 12
Equipment Supplied.......................................................................................................... 12
Equipment Required (Not supplied)....................................................................................... 13
8UNO-COMBINED-CONTROLLER. OVERVIEW.......................................................................................... 13
Temperature Regulation .................................................................................................... 13
CO2Regulation ............................................................................................................... 13
Humidity Regulation.......................................................................................................... 14
9UNO-COMBINED-CONTROLLER. INSTALLATION AND USER GUIDE................................................. 15
Digital Temperature Controller connection ports. ....................................................................... 15
Tubing Connections.......................................................................................................... 16
Pressure Gauge set up and use ........................................................................................... 19
Assembling Instruction of the Bubbling Column......................................................................... 21
Cables connection............................................................................................................ 21
Incubation chamber.......................................................................................................... 24
Fine gauge thermocouple (Free Sensor-optional) ...................................................................... 27
10 CONTROL UNIT FRONT PANEL............................................................................................................ 28
11 TEMPERATURE CONTROL MECHANISM IN STANDARD CONFIGURATION ............................... 29
12 TEMPERATURE CONTROL MECHANISM IN CUSTOM CONFIGURATION AT 37.0°C ................. 31
13 UNIT OPERATION THROUGH PUSH BUTTON INTERFACE............................................................ 34
Main Page, Main Menu And Submenu Description..................................................................... 34
Button functions .............................................................................................................. 35
Main Page and Main Menu Navigation ................................................................................... 36
Changing Set Points......................................................................................................... 37
Selecting Offset Mode....................................................................................................... 37
13.5.1 Standard Offset Mode ................................................................................................ 37
13.5.2 Custom Offset Mode .................................................................................................. 40
Objective Heater Submenu................................................................................................. 41
Humidity Module Submenu................................................................................................. 44
Alarm Settings Submenu.................................................................................................... 45

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Screen Settings Submenu .................................................................................................. 46
Product Information Submenu .......................................................................................... 47
Stop Mode.................................................................................................................. 47
UNO-TS software Active Message..................................................................................... 48
14 OBJECTIVE HEATER MANUAL CALIBRATION.................................................................................. 48
15 FLOW METER REGULATION................................................................................................................. 49
16 MAINTENANCE ........................................................................................................................................ 51
17 SUPPORT.................................................................................................................................................. 52
Web Conference For Assistance And Training.......................................................................... 52
Troubleshooting .............................................................................................................. 53
Technical Support............................................................................................................ 54
ItalyTechnical Specifications ............................................................................................... 54
18 FIGURES.................................................................................................................................................... 55

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1Revision table
Revision Number
Additions or changes
Date
Editor
01
Edited
November 2014
Amelia Liccardo
02
Review
March 2015
Silvia Foppiano
Amelia Liccardo
03
Minor Review
June 2016
Francesco Marino

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2Preface
UNO-COMBINED-CONTROLLER together with a stage top Electric chamber (preventing CO2
of the desired concentration from dissipating away from the sample) is a compact solution for
long term time lapse imaging. We will refer to UNO-COMBINED–CONTROLLER as ‘UNO’ from
here on. ‘UNO’ is designed to maintain all the required thermal and gas conditions for keeping
cells alive for the duration of the time lapse imaging session. “UNO”has a temperature control
range of 25-50°C. The system includes a Temperature Control Unit, a manual Gas Mixer -with
floating ball flow meters-, and a passive Humidity Module with its own heater. The UNO-
COMBINED-CONTROLLER can be connected to a stage top Electric Environmental Chamber
from here on referred to as ‘Incubator’. As OPTIONS a fine gauge thermocouple can be
added to monitor the temperature of a Reference Well located in close proximity to the
experimental one. An Objective Heater is REQUIRED when using oil immersion objectives.
Additionally, UNO-COMBINED-CONTROLLER can be equipped with an optional air pump: OKO-
AP, which is a plug and play solution for Air inlet and a convenient alternative to 100% Air
tanks/compressed Air lines. All the heated components of the ‘Incubator’are heated by 24V
DC and their temperatures monitored by High-Accuracy thermistors. Temperature sensors with
0.1°C accuracy are embedded into the following components: lid and base of the ‘Incubator’,
humidity module and objective-heating collar. A single thermal controller regulates the
temperature of the individual components above in a coordinated fashion avoiding
overshooting the target temperature. Each unit undergoes a careful factory calibration
guaranteeing that the sample temperature, inside the chamber, is kept at the desired value
within the range 25 –50 °C. The system is equipped with visual and sound alarm alert in case
the temperature limit is reached. The unit shows current and set point temperature in the
same display. ‘UNO’ includes an integrated manual CO2mixer allowing regulating Carbon
Dioxide Concentration in Air within the range 1 –20%. Before use please read this manual and
familiarize yourself with the functions and the operation of the device.

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3Symbols Descriptions
3.1.1 Symbols in this manual and on the product:
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 and must refer to
accompanying documents.
Tip Supplies you with helpful suggestions.
Note ►Supplies you with important information to successfully setup and use the instrument.
3.1.2 Symbols on the product label:
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 protection degree against ingress of solids or liquids
inside the product.
4Safety 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.
Technically qualified personnel should only operate equipment.
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
IP 30

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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 rear 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 cables.
Some equipment parts may reach temperatures above 50°C. Please be careful
when handling it.
Check that the cables are well inserted into their connectors so they cannot slip off
This device is not intended for medical use
The display should not be rubbed with abrasive materials or hard objects as it may
be damaged.
Power cord of unit should be unplugged from electrical outlet when left unused for
long period of time.
VENTILATION, please make sure the vents on the unit are not blocked in any way
either partially or fully.
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.
Prevent metal fragments or lead wire scraps from falling inside instrument. This
may cause electric shock, fire or malfunction.
The unit does not contain user serviceable parts.
Before starting, assemble the equipment while unplugged from an outlet.
Unit should never be used where it can fall or be pushed into water.

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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, FINANCIAL OR OTHER LOSSES RESULTING
FROM ANY DEFECTIVE PRODUCT OR THE USE OF ANY PRODUCT.
5AC section wiring diagram
Figure 1 shows the alternating current (AC) section, designed to connect internal power
supplies (see Figure 1 label C) in parallel to low voltage power grid.
Power Connector, OM-BZ0310
Power supplies, TDK-Lambda-ZWS150-24 and ZWS5-5.

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3
1
2L
N
PE
F1
F2
2.5A
2.5A
Power Connector
OM-BZ03105
1.5mm
BLUE
1.5mm
BROWN
24AWG BLACK
24V-DC
24V-DC
0V
0V
0V
0V
1.5mm BLUE
1.5mm BROWN
1mmq
BLUE
1mmq
BROWN
1.5mm
G/V
1mm
BLACK
1mm
RED
5V-DC
0V
Power Supply
TDK-Lambda
ZWS150-24
85 → 265V c.a.
24V c.c.
6.5 A
Power Supply
TDK-Lambda
ZWS5-5
85 → 265V c.a.
5V c.c.
1 A
POSITIVE
CLAMPS
24 V DC
NEGATIVE
CLAMPS
24V-DC
0V
0V
0V
1.5mm G/V
5V-DC
0V
0V
0V
M
24V-DC
0.5mm
BLACK 0.5mm
RED
BLUE
RED
0V
0V
24V-DC
0V
A
B B
Figure 1 AC section wiring diagram

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6UNO-COMBINED-CONTROLLER. Description.
The UNO-COMBINED-CONTROLLER connects Air and CO2 gas (See paragraph 7).
Figure 2. System Overview.
Carbon Dioxide Pressure Gauge with Regulator (Set the pressure at 1bar)
OKO-AP.
Main Control Unit
Humidifying Module with Humidifier Module Heater
Incubating Chamber
Objective Heater (Optional)
When working with Premixed 5% CO2 remember connecting the rigid tube coming
from pressure gauge to Air port labeled “Air In” on rear panel of the Control Unit.
In this working modality, Okolab recommends setting a flow rate of 0.3 L/min using
the floating ball flow meter on the front Panel of the Control Unit.

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7Equipment And Connections
Equipment Supplied
UNO includes:
1. Control-Unit
2. Humidifying Module Heater
3. Glass Bottle - Humidifying Module (attention: FRAGILE) with TUBE E (140 mm
silicon tube of 6 mm OD with Filter).
4. Pressure gauge with filter, regulator and assembly stirrup for CO2, degree of
filtration 20μm and condensate drain. Install pressure gauge between the pure CO2tank
pressure regulator and the Control Unit input port.
5. Power cord cable (x1). Use power cord with Control Unit. The UNO-COMBINED-
CONTROLLER can be powered at a voltage within the range 110-230VAC
6. TUBE E. 3 m long tube assembled as follows: 2 m long rigid tube of 6mm OD,
connected to 0.5m long 4mm ID silicon transparent tube. This assembly is then
connected to an hydrophobic filter 0.2 micron porous and to a 0.5 m long 4mm ID
silicon transparent tube. Connect one end of this tubing assembly to the output port
labeled “Air+CO2 out”and the other end to the Humidity Module.
7. TUBE H. 1.5 m long tube composed of 4mm ID silicon transparent tube (0.5 m long)
connected to 2mm ID silicon transparent tube (1 m long) (both wrapped in an insulating
material). Use insulated tubing in point 8 to connect the Humidity module to the
Incubating chamber.
The following items are required but sold separately.
‘UNO’ must be equipped with:
8. Incubating chamber (model depending on the XY stage with or without z piezo) with
heated lid, chamber riser and one or more plate adapters.
9. Flat screwdriver or Allen wrench. To screw the grub screws or the screws on the
sides of the incubating chamber plugging the built in perfusion holes.
Optional accessories:
OKO-AP + Power cord (x1). The OKO-AP can be powered at a voltage within the
range 110-230VAC
PRESSURE GAUGE WITH FILTER: When a compressed air line is available in the lab
it is possible to avoid using an air pump such as OKO-AP. In this case a Pressure gauge
with filter, regulator and assembly stirrup, degree of filtration 20μm and condensate
drain, for Air is available upon request.
Objective heater. Required ONLY when using oil immersion objectives.

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UNO-TS. UNO-TS software is available for purchase upon request. UNO-TS includes a
Fine Gauge Thermocouple. UNO-TS allows logging temperature data, storing the data in
computer memory as well as performing calibrations in the lab. Additionally, the Fine
Gauge Thermocouple allows operation in Sample Feedback mode.
Equipment Required (Not supplied)
1. CO2gas supply (100%)
2. Compressed Air tank (or a compressed air line). If no air supply is available in the lab
please order air pump OKO-AP.
3. Alternatively to 100% CO2 + air UNO can be operated with a pre-mixed 5% CO2gas
supply.
8UNO-COMBINED-CONTROLLER. Overview
UNO-COMBINED-CONTROLLER is at the same time a temperature and a gas controller. It is
designed to maintain all the required thermal conditions for live cell imaging and is ideal for
time-lapse imaging applications.
Temperature Regulation
UNO-COMBINED-CONTROLLER controls the sample temperature by regulating the
temperature of the base and of the transparent heated lid of the incubating chamber.
Embedded thermistors read the temperature of the incubating chamber’s base, chamber’s lid
and humidifier module. A unique algorithm maintains these temperatures constant. A careful
calibration (See Table 1) performed in our laboratories guarantees that sample temperature is
maintained at the desired value by selecting the Standard Mode (See paragraph 13.5.1)
working modality. An external Fine Gauge Thermocouple (i.e. a thin flexible green wire
temperature sensor (optional) can be used to read the temperature of a reference well to
calibrate the chamber or to check that sample temperature is at the desired value when using
factory default configuration. This optional feature offers higher accuracy of temperature
control. Simply place the thermocouple into the reference well and hold it in place with some
adhesive tape (See paragraph 13.5). The chambers (‘Incubators’) accept different plate
adapters fitting many kinds of plastic supports, such as 35mm Petri Dishes, chambered slides,
simple glass slides and multi-well plates. (See Table 1). Please contact Okolab if you need
assistance selecting the correct incubating chamber fitting your microscope stage brand/model
and sample adapters.
CO2Regulation
CO2is mixed with air in the control unit and is continuously fed into the incubating chamber.
You can also use pre-mixed 5% CO2, connecting the pre-mixed gas supply directly to the
Control Unit, to the input labeled ‘Air In’.

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Humidity Regulation
Humidifying and pre-heating modules prevent medium evaporation and avoid water
condensation on glass and plastic surfaces.

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9UNO-COMBINED-CONTROLLER. Installation And User Guide
The following paragraphs illustrate how to install and use UNO-COMBINED-CONTROLLER.
The following instructions will guide you through a quick installation. For safe
operation of the unit please read carefully all the instructions and safety notes.
Digital Temperature Controller connection ports.
1 2
5 6
3 4
7
9
10
13
14
15
16
8
11
12
Figure 3. Control Unit Rear Panel Overview
1. CO2and Air mixture output
2. RS232 serial port: Software UNO-TS can be used as optional to read incubator
temperature and store the data in computer memory
3. R.T.: Fine gauge thermocouple port
4. Time Lag Fuses 5x20 (x2) (see ‘Power Consumption’in paragraph 17.4)
5. Power Input
6. Power Switch

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7. Heater 1: Chamber Base connector
8. Fuse 1: 1.6 A Fast Fuse
9. Heater 2: Chamber Lid connector
10. Fuse 2: 1.6 A Fast Fuse.
11. H.M.: Humidifying module-heating connector.
12. Fuse 3: 1.6 A Fast Fuse.
13. Obj. Heater: Objective Heater Connector.
14. Fuse 4: 1.6 A Fast Fuse.
15. Pure Air or Premixed Air+5%CO2 Push to fit Input Connector for 6 mm O.D. rigid tubing
16. Pure CO2Push to fit Input Connector for 6 mm O.D. rigid tubing Input
Tubing Connections.
Figure 4. Complete gas lines scheme.

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Figure 5. Complete gas lines scheme. Zoom sketches on the Inlet and Outlet connectors of the Humidifier and Zoom
sketch on the Inlet connector of the Chamber are reported.
1. Your system requires a pressurized CO2 tank (not supplied) with safety valve and
pressure gauge with regulator within the range 0-6 bar.
2. Connect a rigid tube for pressurized gas (not supplied, see Figure 4), with 6 mm Outer
Diameter to the gas output on the pressure regulator of the CO2tank. Alternatively fit
the pressure regulator of the CO2tank with a Swift-Fit Connector for rigid plastic tube
with 6mm O.D. and connect TUBE D (supplied, see paragraph 7.1). If you need further
assistance please contact Okolab.
Secure the Pressure Gauge for CO2with regulator to the wall by using the supplied
assembling stirrup (supplied accessories). A filter is added to the Pressure Gauge.
Filtering degree is 20μm. CO2 stream will be FILTERED and at STABLE Pressure. Do not
fill the filtering container with oil. Connect the rigid tube 6mm O.D., coming from the
pressure regulator of the CO2tank (not supplied), to the Inlet of the Pressure Gauge for
CO2with regulator. Simply push the tubing all the way in. The arrow on the Pressure
Gauge with regulator for CO2indicates the direction of gas flow as illustrated at Figure 7.
3. Please use the rigid tube, TUBE D (supplied, see paragraph 7.1), 3m long 6mm O.D.,
(see Figure 4) to connect the controlled (filtered and at P=1bar) flows of CO2 from the
Pressure Gauge to the Control Unit. Connect TUBE D to the inlet located on the back of

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the controller and labeled “CO2 IN”. Push the tube all the way in into the Swift-Fit
connectors. To remove the tube, pull the tube out while pushing the black ring on the
Swift-Fit connector. Follow the arrow on the Pressure Gauge for the correct gas flow
direction as illustrated in Figure 7. It is important to ensure that the tubing is properly
inserted all the way into the Swift-fit connector in order to avoid gas leaks (See Figure
6).
Figure 6. Avoid gas leak: make sure tubing is pushed all the way into the Push-in to-Fit connector
4. Use the rigid tube, TUBE D (supplied, see paragraph 7.1), to connect the Air Output
Swift-Fit connector on the front panel of the OKO-AP to the Air Inlet Swift-Fit connector
on the rear panel of the Control Unit (Figure 4).
5. Use TUBE E (supplied, see paragraph 7.1) (assembled as follows: 2m long, 6mm O.D.
blue rigid tube + to 0.5 m long 4mm I.D. silicon transparent tube + 0.2 micron porous
hydrophobic filter + 0.5 m long 4mm ID silicon transparent tube) to connect the Air +
CO2 output Swift-Fit connector on the rear panel of the Control Unit to the Inlet barb
connector on the Humidifier bottle, as illustrated in Figure 4 and Figure 5.
6. Use TUBE H (supplied, see paragraph 7.1) (assembled as follows: 0.5 m long 4mm ID
silicon transparent tube + 1 m long, 2mm ID silicon transparent tube wrapped in
insulating material) to connect the Outlet barb connector on the Humidifier bottle to the
miniature barb connector on the Incubating Chamber, as illustrated in the Figure 4 and
in Figure 5.
It is possible to use additional filters or filtering devices to further filter the gas mixture
prior to its entrance into the incubation chamber further reducing the risk of
contamination. If you choose to install an additional filter insert it after the Control Unit
and before the hydrophobic filter included in the tubing assembly of TUBE E.
Note Use OKO AP when no Air pressurized tank is available.
Note Whenever your system is equipped with Premixed gas pressurized tanks with
safety valve and pressure gauge with regulator within the range 0-6 bar (not supplied),
you can directly connect the Premixed gas to the Air inlet of the Control Unit using Tube
D and use the Air regulation in the front panel to control the flow.

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Do not remove the hydrophobic filter along the tubing, TUBE C, which connects the
Air + CO2output Swift-Fit connector on the rear panel of the Control Unit to the
Inlet barb connector on the Humidifier bottle. Replace that filter every 6 months.
Contact Okolab for further information.
Avoid pinching and kinking of all tubing as it will hinder gas flow and may cause
condensation. It is recommended to periodically check tubing connections and gas
flow.
Pressure Gauge set up and use
Follow the direction of arrow on the rear of the Pressure Gauge for the correct Gas In-Out (See
Figure 7 below).
GAS IN GAS OUT
Figure 7.Input and Output gas ports. See symbol on the rear of Pressure Gauge.
On the bottom of the unit there is a purging valve. Close it before operating the Pressure
Gauge. To close the purging valve -looking at the regulator vertically from the top knob- rotate
it counterclockwise. (See A and B in Figure 8). Now you can operate the Pressure Gauge.

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Purging valve
A B
Figure 8. How to close the purging valve of the Pressure Gauge.
To close the gauge, pull the knob up and rotate it counterclockwise. Push the knob down to
lock it. Open valve on gas supply tank. Adjust Pressure Gauge by pulling the knob up and
rotating it clockwise until the pressure reaches the desired value (See Figure 9).
A B C
Figure 9. Pressure gauge using.