UNO STAGE TOP INCUBATOR All In One Stage Top Incubator. Manual

UNO STAGE TOP INCUBATOR All In One Stage Top Incubator Manual Vers. 3-2017 1

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Index Index PREFACE... 5 2 SYMBOLS DESCRIPTION... 6 SYMBOLS IN THIS MANUAL AND ON THE PRODUCT... 6 SYMBOLS ON THE PRODUCT LABEL... 6 3 SAFETY NOTES... 7 4 SYSTEM OVERVIEW... 9 UNO CONTROL UNIT... 9 4.1.1 BUNDLE 1) UNO-T. Overview and supplied equipment... 11 4.1.2 BUNDLE 2) UNO-T-H-PREMIXED. Overview and supplied equipment... 12 4.1.3 BUNDLE 3) UNO-T-H-CO2. Overview and supplied equipment... 13 STAGE TOP CHAMBER... 15 5 INSTALLATION GUIDE... 15 ASSEMBLY AND GAS CONNECTION OF THE BUBBLING COLUMN... 15 CONNECTIONS OF THE HEATING DEVICES... 16 5.2.1 Connection of the Chamber and of the Lid of the Incubating Chamber... 16 5.2.2 Connection of the Humidifier and of the Heated Tube... 17 5.2.3 Connection of the Objective Heater (optional)... 18 CONNECTION OF THE GAS TUBES... 18 5.3.1 Gas connection for UNO-T-H-PREMIXED... 18 5.3.1.1 How to use the GF Regulator... 19 5.3.2 Gas Connection for UNO-T-H-CO2... 20 5.3.2.1 How to use the Air/CO2 Mixer... 20 CONNECTION OF THE TEMPERATURE SENSOR... 22 MINI-USB PORT... 23 5.5.1 Connection of a USB drive... 23 5.5.2 PC Connection via USB... 23 RS232 PORT PC CONNECTION VIA RS232... 24 INSTALLATION OF THE STAGE TOP CHAMBER... 24 6 USER INTERFACE... 25 HOMEPAGE... 25 6.1.1 Colors meaning... 26 6.1.2 Changing the set point temperature... 26 6.1.3 Graph view... 26 SYSTEM INFORMATION... 27 SETTINGS... 27 6.3.1 Control Mode... 28 6.3.1.1 Open Incubator... 30 6.3.2 Chamber & Insert... 31 6.3.2.1 Compatible chambers... 32 6.3.3 Objective Heater... 34 6.3.4 Humidity... 35 6.3.5 Calibration... 36 6.3.5.1 Chamber Calibration... 36 6.3.5.2 T sensor Calibration... 37 6.3.5.3 Objective Heater Calibration... 39 6.3.6 Logging... 40 6.3.7 Alarms... 42 6.3.8 Display... 44 6.3.8.1 Options... 44 6.3.8.2 Brightness... 45 6.3.8.3 Calibration... 45 3

6.3.8.4 Date & Time... 45 6.3.8.5 Visual Effects... 46 6.3.8.6 Factory Reset... 46 6.3.9 Password... 47 6.3.10 Summary... 48 6.3.11 Auto on/off... 48 STATUS... 49 GLANCE MODE... 50 7 MAINTENANCE... 51 8 SUPPORT... 52 SUPPORT PAGE... 52 SUPPORT CONTACTS... 53 WEB CONFERENCE FOR ASSISTANCE AND TRAINING... 53 9 TECHNICAL SPECIFICATIONS... 54 10 TROUBLESHOOTING... 55 11 FIGURE LIST... 56 12 TABLE LIST... 57 13 MANUAL REVISION TABLE... 58 14 LIST OF SPARE PARTS... 59 UNO-T-H-PREMIXED... 59 UNO-T-H-CO2... 59 4

Preface UNO STAGE TOP INCUBATOR is a Digital Temperature (and Humidity) Controller and a data log device enabling the control of the environment inside the top stage incubator and is composed by UNO CONTROL UNIT and a STAGE TOP CHAMBER. UNO CONTROL UNIT ensures the highest temperature accuracy on sample temperature by controlling simultaneously up to 6 independent channels with a modern multichannel controller. UNO CONTROL UNIT comes in three different setups: UNO-T UNO CONTROL UNIT (only) UNO-T-H-PREMIXED UNO CONTROL UNIT Humidity module Flow rate regulator for premixed gas UNO-T-H-CO2 UNO CONTROL UNIT Humidity module CO2/Air gas mixer Air Pump The OBJECTIVE HEATER is an optional device which can be purchased upon request. It ensures the absence of a heat sink when the microscope objective is in close contact with the specimen. Please contact Okolab for more information on this device. The end user can define the tolerance values, which can trigger an alarm. UNO CONTROL UNIT allows you also to log your experimental data via USB connection (or RS232 connection), using an OKOLAB software, DATALOG (purchased on request) or a third party software. Moreover, UNO CONTROL UNIT can store data on its internal memory and these data can be then downloaded on a USB stick at user request. Reading this manual is essential to familiarize yourself with the installation, the functions and the operation of the device before use. 5

2 SYMBOLS DESCRIPTION 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. Please refer to accompanying documents. Tip - Supplies you with helpful suggestions. Note - Supplies you with important information to successfully setup and use the instrument. 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 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 6

3 SAFETY 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 be used only as intended and described in this Manual. Equipment should be operated only 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 subject to flammable or explosive gases. Transport the equipment with care. The 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 damage or breakdown the device and its accessories. The product labels can be found on the bottom panel of the Main Unit. Do NOT make in close contact with or breathe any gas stream whose composition is different from that of ambient air. Do not exceed the voltage indicated in this manual and on the product label. Avoid excessive induction noise, static electricity, magnetic fields. Do not expose this instrument to water or moisture. Prevent throttling and kinking of tubing. Check tubing from time to time for possible wear and tear. Check that all tubing are well inserted into connectors so they cannot slip off. This device has not been designed for use for medical applications. Install safety valves and adequate pressure regulators on gas lines before the Unit input connectors. The power cord of the unit should be unplugged from the electrical outlet when left unused for a long time period. PRESSURIZED GAS, secure all connections with hose clamps. Never exceed pressure limits, 1.8 bar for N 2 and 5 bar for CO 2. Bleed all lines before disconnecting. Wear safety goggles if needed at all the time. If the pressure regulators are not within easy sight and reach make sure there is at least one shut off valve for each line. LOW OXYGEN ATMOSPHERES and HIGH CO 2 ATMOSPHERES, never enter a room or enclosure which has a low oxygen atmosphere or a high CO 2 atmosphere because of the severe danger of suffocation. Only operate in a well-ventilated room. A small amount of gas mixture (nitrogen and carbon dioxide) leaks continuously out of the instrument and should never be allowed to build up in the room. 7

VENTILATION, the unit should be placed so that its location or position does not interfere with proper ventilation. Neither the gas controller nor the stream destinations should be in poorly ventilated areas. The unit should be placed far from heat sources such as open flames, 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 a grounded mains power socket. Do not disconnect cables while in operation. Prevent metal fragments or lead wire scraps from falling inside the instrument case to avoid electric shock, fire or malfunction. Before starting, assemble the equipment far from any electrical devices. The unit should never be used where it can fall or be pushed into water. International caution symbol marks this device. It is important to 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, OR LOSSES OR EXPENSES RESULTING FROM ANY DEFECTIVE PRODUCT OR THE USE OF ANY PRODUCT. 8

4 SYSTEM OVERVIEW This Chapter describes the provided equipment. For your UNO STAGE TOP INCUBATOR that comes in three different bundles: Bundle 1) UNO-T. Stage Top Incubator for application where only thermal control is required (no gas/no humidity), which includes (see paragraph 4.1.1): a. UNO CONTROL UNIT b. Stage Top Chamber Bundle 2) UNO-T-H-PREMIXED. Stage Top Incubator for application with pre-mixed gas, which includes (see paragraph 4.1.2): a. UNO CONTROL UNIT b. Humidity module c. Flow rate regulator for premixed gas d. Stage Top Chamber Bundle 3) UNO-T-H-CO2. Stage Top Incubator for application with 100% CO2/Air mixer, for application with 100% CO2 and background Air, which includes (see paragraph 4.1.3): a. UNO CONTROL UNIT b. Humidity module c. CO2/Air gas mixer d. Air Pump e. Stage Top Chamber UNO CONTROL UNIT Figure 1 shows the UNO CONTROL UNIT overview; with reference to the highlighted pointers: 1. MINI-USB port. It allows to connect the UNO CONTROL UNIT to a PC via USB or to a USB stick, see paragraph 5.5. 2. RS 232 port. It allows to connect the UNO CONTROL UNIT to a PC via USB or to a USB drive, see paragraph 5.6. 3. Power Input. 4. Base/lid. It is the power outlet for the Chamber s Base and Lid, see paragraph 5.2.1. 5. Humidifier/Objective Heater. It is the power outlet for the Humidifier/Bubbling Column and the Objective Heater(optional), see paragraph 5.2.2. 6. Heated Tube. It is the power outlet for the Heated Tube, see paragraph 0. 7. Temperature Sensor port. It allows to connect the Temperature Sensor to UNO CONTROL UNIT, see paragraph 5.3. 8. ON/OFF button. 9

Figure 1. UNO CONTROL UNIT overview. 10

4.1.1 BUNDLE 1) UNO-T. Overview and supplied equipment The equipment that comes with UNO-T is composed by: 1. T-Sensor 2. UNO CONTROL UNIT (see paragraph 4.1) 3. Y-CABLE-1 (see paragraph 5.2.1) 4. Objective Heater (optional) 5. STAGE TOP CHAMBER (see paragraph 4.2) 6. Power adapter (voltage: 24V; max power consumption: 110W; max current: 4.6A) 7. MINI-USB-OTG (see paragraph 5.5) Figure 2. UNO-T. 11

4.1.2 BUNDLE 2) UNO-T-H-PREMIXED. Overview and supplied equipment The equipment that comes with UNO-T-H-PREMIXED is composed by: 1. T-Sensor 2. UNO CONTROL UNIT (see paragraph 4.1) 3. GF-REGULATOR (see paragraph 5.3.1 and 5.3.1.1) 4. TUBE-C (see Table 1) 5. Humidifier 6. Heated Tube (see paragraph 0) 7. Objective Heater (optional) 8. STAGE TOP CHAMBER 9. Y-CABLE-1 (see paragraph 5.2.1) 10. Y-CABLE-2 (see paragraph 5.2.2) 11. Power adapter (voltage: 24V; max power consumption: 110W; max current: 4.6A) 12. MINI-USB-OTG (see paragraph 5.5) Figure 3. UNO-T-H-PREMIXED. Description Photo TUBE-C (X1) Composed tube 300 cm long: Transparent PU O.D. 6mm I.D. 4mm, Length 200 cm + Plastic Connector Promepla 4,8-3,2. + Sil O.D. 7mm I.D. 4mm, Length 50 cm + PTFE 0.2 micron + Sil O.D. 7mm I.D. 4mm, Length 50 cm Table 1. Tubes for UNO-T-H-PREMIXED. 12

4.1.3 BUNDLE 3) UNO-T-H-CO2. Overview and supplied equipment The equipment that comes with UNO-T-H-CO2 is composed by: 1. OKO-AP (Air Pump) (see paragraph 5.3.2 and 5.3.2.1) 2. TUBE-A (see Table 2) 3. 2GF-MIXER (Gas Manual Mixer to mix Air and CO2) (see paragraph 5.3.2 and 5.3.2.1) 4. Pressure Gauge (see paragraph 5.3.2) 5. TUBE-A (see Table 2) 6. TUBE-C (see Table 2) 7. Glass Bottle - Humidifying Module (attention: FRAGILE) 8. Heated Tube (see paragraph 0) 9. Objective Heater (optional) 10. STAGE TOP CHAMBER 11. Y-CABLE-1 (see paragraph 5.2.1) 12. Y-CABLE-2 (see paragraph 5.2.2) 13. Power adapter (voltage: 24V; max power consumption: 110W; max current: 4.6A) 14. UNO CONTROL UNIT (see paragraph 4.1) 15. MINI-USB-OTG (see paragraph 5.5) 16. T-Sensor Figure 4. UNO-T-H-CO2. 13

Description Tube-A (X2) Single Tube 300 cm long: Transparent PU O.D. 6 mm I.D. 4mm, Length 300 cm. Photo TUBE-C (X1) Composed tube 300 cm long: Transparent PU O.D. 6mm I.D. 4mm, Length 200 cm + Plastic Connector Promepla 4,8-3,2. + Sil O.D. 7mm I.D. 4mm, Length 50 cm + PTFE 0.2 micron + Sil O.D. 7mm I.D. 4mm, Length 50 cm Table 2. Tubes for UNO-T-H-CO2. 14

STAGE TOP CHAMBER For the equipment description of the Stage Top Chamber, refer to the dedicated attached Stage Top Chamber manual. 5 INSTALLATION GUIDE Assembly and Gas connection of the Bubbling Column The bubbling column comes with UNO-T-H-PREMIXED and UNO-T-H-CO2; follow the instructions below to correctly assemble it (see Figure 5): 1. Unscrew the red cap with O-ring and the head of the bubbling column from the bottle, without removing the head from the plug. 2. Insert filter into the silicone tubing. Connect the silicone tubing to port of the head of the bubbling column on the inside of the red screw cap. Figure 5. Assembly instruction of the Bubbling Column and its parts. Follow the instructions below to correctly connect the gas tubes to the Bubbling Column (see Figure 6): 1. Plug the silicon end of TUBE-C to the Bubbling Column inlet (metal plug). 2. Push the Heated Tube from the PU end (yellow tube) all the way in to the push-to-fit Bubbling column output. 3. Secure the Heated Tube silicon end to the chamber, making sure the tube is all the way in. 15

Figure 6. Gas connections of the bubbling column. Connections of the heating devices This section describes how to connect the heating devices to UNO CONTROL UNIT. 5.2.1 Connection of the Chamber and of the Lid of the Incubating Chamber Follow the instructions below to connect the Chamber and the Lid of the Incubating Chamber to UNO CONTROL UNIT (see Figure 7): 1. Connect the Y-CABLE-1 to power output of Base and Lid on the back panel of UNO CONTROL UNIT. The power output for the Chamber and for the Lid is the one on the left, labeled Base/Lid; 2. Connect the Chamber electric cable to its connector on the Y-CABLE-1 (Chamber color label is red); 3. Connect the Lid electric cable to its connector on the Y-CABLE-1 (Lid color label is yellow). Figure 7. Chamber and Lid of the Incubating Chamber connection. 16

5.2.2 Connection of the Humidifier and of the Heated Tube Follow the instructions below to connect the Humidifier and the Heated Tube of the Incubating Chamber to UNO CONTROL UNIT (see Figure 8): 1. Connect the Y-CABLE-2 to power output of Humidifier and Heated Tube on the back panel of UNO CONTROL UNIT. The power output for the Humidifier and for the Heated Tube is the one in the middle, labeled Humidifier/Heated Tube; 2. Connect the Humidifier cable to its connector on the Y-CABLE-2 (Humidifier color label is green); 3. Connect the Heated Tube cable to its connector on the Y-CABLE-2 (Heated Tube color label is gray). Figure 8. Humidifier and Heated Tube connection. 17

5.2.3 Connection of the Objective Heater (optional) To connect the Heater Tube to UNO CONTROL UNIT, connect the Objective Heater on the back panel of UNO CONTROL UNIT. The power output for the Objective Heater is the one on the right, labeled Objective Heater (see Figure 9). Figure 9. Objective Heater connection. The cables connecting heating devices are not only color coded, but there is also a label glued to their end. Connection of the gas tubes 5.3.1 Gas connection for UNO-T-H-PREMIXED UNO-T-H-PREMIXED works for application with pre-mixed gas. To make the gas flow into the chamber the steps are the following: 1. Connect your Premixed Gas tank to the supplied GF Regulator inlet (During this operation make sure that both the tank and the GF Regulator are closed). 2. Connect the GF Regulator output to the Humidifier bottle using TUBE-C. 18

Figure 10. UNO-T-H-PREMIXED gas connection. 5.3.1.1 How to use the GF Regulator The GF regulator allows to deliver the exact amount of gas flow, in fact since it is provided with a calibrated orifice, it is possible to control the pressure in order to have the desired gas flow. Table 3 shows the relation between Pressure and Flow for the GF Regulator. Pressure [barg] Flow [l/min] 0,2 0,07 0,4 0,10 0,6 0,12 0,8 0,14 1,0 0,17 1,2 0,18 1,4 0,20 1,6 0,22 1,8 0,23 2,0 0,25 2,2 0,27 2,4 0,29 2,6 0,31 2,8 0,32 3,0 0,34 3,2 0,36 3,4 0,38 3,6 0,39 3,8 0,4 Table 3. UNO-T-H-PREMIXED Pressure/Flow relation. Example: the desired gas flow is 0,2, hence the GF Regulator has to be set to 1,4 barg. 19

5.3.2 Gas Connection for UNO-T-H-CO2 UNO-T-H-CO2 includes the Air/CO2 mixer and the OKO-AP, it works for application with 100% CO2 and background Air. To make the gas flow into the chamber the steps are the following: 1. Connect your CO2 Gas line/tank to the supplied Pressure Gauge inlet (During this operation make sure that both the tank and the GF Regulator are closed). The CO2 entering the Pressure Gauge has to be set at 2 bar. 2. Connect the Pressure gauge to the CO2 In, which is located on the left of the rear panel of the Air/CO2 Mixer using TUBE-A. 3. Connect the OKO-AP to the Air In, which is located on the right of the rear panel of the Air/CO2 Mixer using TUBE-A. 4. Connect the Air+CO2 OUT, located on the rear panel of the Air/CO2 Mixer, to the Humidifier bottle using TUBE-C. Figure 11. UNO-T-H-CO2 gas connection. 5.3.2.1 How to use the Air/CO2 Mixer The Air/CO2 Mixer allows to generate CO2-Air mixtures with an adjustable CO2 concentration in the range 0-15%. Air and CO2 flows are regulated by two floating ball flow meters in the range 0.1 1 Nl/min and 0.013 0.13 Nl/min, respectively. As an example, Table 1 reports a set of flow rates to obtain a concentration of CO2 equal to 5%. A more comprehensive table is reported in the manual. Do not use the flow meters as closing-opening valves. Once regulated the CO2 flow pressure (at P= 1bar, see Figure 11), use the flow meters carefully only for fine regulations. 1. Open the gas valves about 15 minutes before inserting the sample in the Micro Environmental Chamber. Please follow carefully scheme in Figure 11. 20

Target Gas Percentage [%] Gas flow rate values to set [Nl/min] Air CO2 Air CO2 Total 95.00 5.00 0.60 0.03 0.63 95.00 5.00 0.80 0.04 0.84 95.00 5.00 1.00 0.05 1.05 Table 4. Flow meter regulation, two flow meters control unit. [Nl = Normal litre] 2. Target Gas Percentage indicates the stream gas percentage that the operator wants during the experiment. 3. Gas flow rate values to set, in Normal litre per minute [NL/min], are the values of gas flow rate to set on the graduated scale (see Figure 12). 4. Total indicates the measured total gas stream flow rate. Tip 1. If you need, ask Okolab for personalized Gas Percentage Tables Example: the target Air percentage is 95.00%, the target CO 2 percentage is 5.00%. In Table 4 three possibilities that realize the required gas percentage are. Notice that the values are a compromise between CO 2 consumption, condensation (normally it decreases if Total gas stream flow rate increases) and medium evaporation (normally it decreases if Total gas stream flow rate decreases). We could consider the second line in Table 4. So, we regulate the two flow meters to obtain the desired relative quantity of air and CO 2, as shown in Figure 12. Notice that the correct position of the indicator (ball) is about in its middle. Figure 12. Flow meters graduated scales, example for two gas lines. Note 1. If you use a single reservoir of air+5% CO 2 the total gas steam flow rate suggested is 0.1 L/min. 21

Connection of the Temperature Sensor Connect the Temperature Sensor to the real panel, precisely to the green housing on the right, as indicated in Figure 13. Figure 13. Temperature Sensor connection. Tip 1. The Temperature Sensor has to be connected with + on top. 22

MINI-USB port The MINI-USB port allows the UNO CONTROL UNIT to be connected with the outside world. 5.5.1 Connection of a USB drive Use MINI-USB-OTG cable to connect a USB Drive (not supplied) to the CONTROLLER, as shown in Figure 14. Figure 14. USB drive connection. 5.5.2 PC Connection via USB Use a standard MINI-USB cable to connect UNO CONTROL UNIT to a PC, as shown in Figure 15. You can communicate with UNO CONTROL UNIT via PC by using: DATA-LOG software. The software allows to log and analyze the data provided by UNO CONTROL UNIT via PC. Refer to DATA-LOG user manual for more info. SDK (OKOLAB API) for integration in third party software http://www.okolab.com/support#soft_int_tools. Figure 15. PC connection via USB. 23

RS232 port PC connection via RS232 Use a standard RS232 cable to connect UNO CONTROL UNIT to a PC as shown in Figure 16. This option is advisable when the PC is at a distance greater than 10 meters from the CONTROLLER. You can interface with UNO CONTROL UNIT via PC by using: DATA-LOG software. The software allows to log and analyze the data provided by CONTROLLER via PC. Refer to DATA-LOG user manual for more info. SDK (OKOLAB API) for integration in third party software http://www.okolab.com/support#soft_int_tools. Figure 16. PC connection via RS232. Without a serial port COM, to install the Serial Communication cable to your PC, you need to install a USB to serial converter following the manufacturer instructions. If you want to log, you have to use the COM port number that Windows assigned to the drive. Installation of the Stage Top Chamber For the installation of the Stage Top Chamber refer to the dedicated attached Stage Top Chamber manual. 24

6 USER INTERFACE This chapter describes the user interface of the UNO CONTROL UNIT. Homepage Figure 17 shows the main control panel, in particular it highlights 19 pointers detailed below. Figure 17. Home page. 1. System Information. Press here to know generic info about UNO and its running time. See paragraph 6.2. 2. Home. Press here to return to the home page. 3. Settings. Press here to access system options and settings. See paragraph 6.3. 4. Status. Press here to see the status of your system (you will need this page in case of assistance). See paragraph 6.4. 5. Glance mode. Press here to switch the display to glance mode. See paragraph 6.3.8.5 and 6.5. 6. UNO status led. See paragraph 6.1.1 for the color mapping. 7. Chamber (or T Sensor) current temperature. 8. Temperature set point. 9. Minimum and Maximum temperature values in the actual time window. 10. Temperature at the heating tube. 11. Temperature at the T sensor. 12. Temperature at the lid. 13. Temperature at the base. 14. Temperature at the Objective heater. 15. Support. Press here to learn how to contact OKOLAB. See paragraph 8.1. 16. Temperature at the Bubbling Column. 17. Open Incubator. 18. PC or USB Connection indicator. 19. Logging icon. 20. Temperature evolution graph. 25

6.1.1 Colors meaning The GREEN indicator indicates that the set-point value has been reached (within the tolerance you have set in the alarm subpage, see paragraph 6.3.7) and that the system is working properly. The YELLOW indicator means that the system is working towards reaching the set-points (transient state). No actions on your part are required. Please note the Yellow light will also appear every time you change temperature set point. The ORANGE indicator means that the current temperature is out of the tolerance you have set (see paragraph 6.3.7). Verify that all cables are properly connected. Check all the enclosures to verify that they are properly closed. The RED indicator means that there is a problem with the unit itself (for example the Temperature sensor has been damaged). Turn the system off, wait 5 minutes, and turn it back on. If the problem continues, contact Okolab at www.oko-lab.com 6.1.2 Changing the set point temperature To input a new set-point temperature for a heating device, touch the corresponding tab, indicated as Pointer 1 in Figure 18(a). The set point regulation page will appear as in the Figure 18. You can modify the set point by clicking on +/-, see Pointer 2. Once you have input the new set point temperature, press Set to save (or Cancel to undo), see Pointer 3. Tip 2. After any change in the set point value the controller enters into a transient regime, the Status Indicator and the Top LED turn yellow (see paragraph 6.1.1). During the transient regime, the controller will not trigger on alarm. The maximum length of the transient regime is set by the operator as indicated in paragraph 6.3.7. (a) Figure 18. Changing the set point temperature. 6.1.3 Graph view By pressing on the Temperature evolution chart (pointer 1 in Figure 19(a)), the Graph view page will open. By checking or unchecking the checkbox Grid (pointer 2 in Figure 19), it is possible to see or to remove, respectively, the grid behind the graph. 26

(a) Figure 19. Graph view. System Information Press the icon related to the Controller version. to access the Info Page, as shown in Figure 20. This page contains the information Figure 20. Info page selection Tip 3. Please have this information handy when contacting Okolab for support. Settings Press on Settings icon to enter the Settings menu, as shown in Figure 21. Figure 21.How to access the Settings page. 27

The settings menu has 11 items, as Figure 22(a) and Figure 22 show: 1. Control Mode. See paragraph 6.3.1. 7. Alarms. See paragraph 6.3.7. 2. Chamber & Insert. See paragraph 6.3.2. 3. Obj. Heater. See paragraph 6.3.3. 4. Humidity. See paragraph 6.3.4. 5. Calibrations. See paragraph 6.3.5 8. Display. See paragraph 6.3.8. 9. Password. See paragraph 6.3.9. 10. Summary. See paragraph 6.3.10. 11. Auto on/off. See paragraph 6.3.11. 6. Logging. See paragraph 6.3.6. (a) Figure 22. Graph view. Each of the aforementioned item will be detailed in the following. 6.3.1 Control Mode Press on to enter the Control Mode menu, as Figure 23 shows. Figure 23. Control Mode menu. Once you are in the Control Mode submenu it is possible to choose: 1. Sample Control mode. Figure 24 shows how to select Sample Control mode: Press on the Sample icon; Press Save. 28

Now the UNO CONTROL UNIT will start operating in Sample Control Mode. Figure 24. Sample Control mode. Note 1. Sample Control mode allows you to take the temperature feedback from the specimen with the dedicated Temperature Sensor. This mode gives the opportunity to control the environment directly inside the plate with the drawback of having a plate dedicated to the temperature acquisition. 2. Chamber Control mode. Figure 25 shows how to select Chamber Control mode: Press on the Chamber icon; Press Save. Figure 25. Chamber Control mode. Note 2. Chamber Control mode allows you to take the temperature feedback directly from the chamber temperature sensor, without the need of having floating wires for a dedicated specimen s Temperature Sensor. At this point the page Chamber Offset will pop out, see Figure 26. In fact, there are two options for operating in Chamber Control mode: 1. Factory: by pressing on the Factory button the UNO CONTROL UNIT will use the calibration parameters already optimized by Okolab. 2. Calibrate: by pressing on the Calibrate button the UNO CONTROL UNIT will start a routine to calibrate its parameters with the help of the external Temperature Sensor, to be placed inside the chamber before the operation. Figure 27(a) shows the UNO CONTROL UNIT during the calibration procedure; by clicking Options, the menu Advanced calibration parameters will appear and, here, it is possible to set the calibration Accuracy and Precision, see Figure 27. 29

Figure 26. Chamber Control mode Chamber Offset. (a) Figure 27. Chamber Control mode Self Calibration. 6.3.1.1 Open Incubator During the normal operation it could be necessary to open the incubator. Figure 28 shows the procedure to follow when opening incubator: 1. Press on Open Incubator, before removing the lid, as Figure 28 (a) shows. Now you can open the incubator; 2. Once you are done close the chamber and wait for 2 minutes in order to let the environment achieve steady state; press on resume operating normally. Incubator Closed, as Figure 28 shows, and the UNO CONTROL UNIT will (a) Figure 28. Control mode Open Incubator. 30

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, UNO CONTROL UNIT 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. 6.3.2 Chamber & Insert UNO CONTROL UNIT is available for a plethora of chambers to perfectly fit your specific microscope setup and the plates you need to use. Since the bottom of the chamber is in contact with the external environment, the UNO CONTROL UNIT has to modify its controlling parameters in dependence of the specific couple Chamber/Insert. To setup your Chamber/Insert configuration press on Chamber & Insert, as Figure 29(a) shows. At this point the Chamber & Insert submenu in Figure 29 will appear. (a) Figure 29. Chamber & Insert. Figure 30(a) and show the selection menus for Chamber and Insert, respectively. You can navigate through the menus to choose the objects present in your UNO CONTROL UNIT configuration. (a) Figure 30. Chamber & Insert configuration. (a)chamber selection; Insert selection. 31

6.3.2.1 Compatible chambers Table 5 reports all the available chambers for UNO STAGE TOP INCUBATOR. Chambers: H301-K-FRAME H301-NIKON-TI-S-ER H301-OLYMPUS-IX3-SVR H301-PRIOR-H117 Detail: Fits standard XY stages with k-frame insert (160x110mm). Chambers: H301-LUDL96A602 H301-MCL-Z100/500 H301-NIKON-NZ100/200/500-N H301-PI-736-160x110 Detail: Fits Z-piezo stages models listed. Chambers: H301-MINI Detail: Fits in any MW plate holder. Chambers: H301-NIKON-TI-SR H301-OLYMPUS-IX-SUSP Detail: Fits rectangular stages with circular opening. Chambers: H301-EC-LG-BL Detail: Fits Leica Super Z Galvo Stage (SP5/SP8). Chambers: H301-PRIOR-NZ500 Detail: Fits Prior Z-piezo stages models NZ250/400/500. 32

Chambers: H301-PRIOR-NZ100-H107 H301-PRIOR-NZ100-H117 H301-PI-736-ZR15/ZR25 Detail: Fits Prior Z-piezo stages models listed. Chambers: H301-UP Detail: Fits any Upright Microscopes. It allows the usage of dipping lenses. Chambers: H301-EC-LG-UP-BL Detail: Fits Upright Leica Super Z Galvo Stage. It allows the usage of dipping lenses. Chambers: H301-DUAL-GAS Detail: Chamber with two independent gas compartments. Chambers: CUSTOM Detail: Parameters for custom chambers. Table 5. Compatible chambers. 33

6.3.3 Objective Heater Press on Objective Heater icon to enter the Objective Heater menu, see Figure 31(a). The Objective Heater is disabled by default, to activate it press on the Enabled icon, see Figure 31. At this point you can choose: 1. Ramp Rate. By pressing on this button the submenu in Figure 32 will appear; here you can tune the ramp rate for the temperature of the Objective Heater. 2. Cancel. To Cancel your choice. 3. OK. To confirm your choice; moreover, after pressing this button, the menu in Figure 33 will appear giving you further options. (a) Figure 31. Objective Heater. (a) How to access the Objective Heater menu; Objective Heater menu. Figure 32. Ramp rate. Before enabling the Objective Heater, make sure that it is correctly mounted around your objective and that it is connected to the UNO CONTROL UNIT. Figure 33 shows the menu that appears when selecting the Objective Heater feature, with reference to the Pointers 4 options are available: 1. Log: by pressing this button you can recall parameters obtained with previously performed calibrations, see Figure 34(a); 2. Factory: by pressing this button UNO CONTROL UNIT will resume the factory offset; 3. Manual: by pressing this button can find the best offset value manually, step by step, see Figure 34; 4. Calibrate: by pressing this button UNO CONTROL UNIT will perform a calibration routine for your specific environment, see paragraph 6.3.5. 34

Figure 33. Objective Heater Enabled. (a) Figure 34. Objective Heater options. (a) Log; Manual. 6.3.4 Humidity Press on Humidity icon to enter the Humidity menu, see Figure 35. Figure 35. How to access the Humidity menu. Figure 36(a) shows the Humidity menu. Once the Humidity control enabled checkbox is checked, UNO CONTROL UNIT will perform humidity control acting on the temperatures of both the humidifier s bottle and the heated tube. In order to make the humidity control work properly, the user has to set the actual room temperature, see Figure 36(a). UNO CONTROL UNIT, by default, supplies a humidity level of 90%; whereas when you require a different humidity level, it is possible to act on the Offset temperature, see Figure 36: rising this value will lead to rising the humidity level as well. 35

(a) Figure 36. Humidity menu. (a) Main menu; Offset. 6.3.5 Calibration Press on Calibration icon to enter the Settings menu, as shown in Figure 37. Figure 37. How to access Calibration menu. Once you are in the Control Mode submenu it is possible to choose: 1. Chamber Calib. Here you can perform the calibration of the chamber, see paragraph 6.3.5.1. 2. T Sensor. Here you can perform the calibration of the Temperature Sensor, see paragraph 6.3.5.2. 3. Obj calibration. Here you can perform the calibration of the Objective Heater, see paragraph 6.3.5.3. 6.3.5.1 Chamber Calibration Press on Chamber Calib icon 39., as Figure 38 shows, to enter the Chamber Calibration submenu in Figure Figure 38. Chamber calibration. 36

Two options are available to the Chamber Calibration, see Figure 39: 1. Factory: by pressing on the Factory button the UNO CONTROL UNIT will use the calibration parameters already optimized by Okolab. 2. Calibrate: by pressing on the Calibrate button the UNO CONTROL UNIT will start a routine to calibrate its parameters with the help of the external Temperature Sensor, to be placed inside the chamber before the operation. Figure 40(a) shows the UNO CONTROL UNIT during the calibration procedure; by clicking Options, the menu Advanced calibration parameters will appear and, here, it is possible to set the calibration Accuracy and Precision, see Figure 40. Figure 39. Chamber Control mode Chamber Offset. (a) Figure 40. Chamber Control mode Self Calibration. 6.3.5.2 T sensor Calibration The Temperature Sensor of UNO CONTROL UNIT is calibrated in Okolab at three different temperatures, i.e. 35.0, 37.0 and 40.0 C. To view the factory calibration data, press on the T sensor calibration icon as shown in Figure 41(a) and then on the tab labeled Factory as shown in Figure 41; the page with the calibration data will open, as shown in Figure 42. 37

(a) Figure 41. Calibration data. (a) T sensor calibration page selection; T sensor calibration page. Figure 42. Free sensor factory offset To re-calibrate the T sensor, follow the instructions below. 1. Set a water bath to the temperature you use as set point for the heating devices during standard operation (most of the times it is 37 C); 2. Measure water temperature by immersing the Temperature Sensor and a Certified Thermometer; 3. Type in the reading of the certified thermometer by pressing on the pencil, as shown in Figure 43(a); 4. Press on Set to adjust to reading of the Temperature Sensor against the reading of the certified thermometer, as shown in Figure 43. (a) Figure 43. How to re-calibrate the T sensor Tip 4. Press on icon to receive instructions on how to calibrate the T-Sensor. Tip 5. Okolab recommends using a certified thermometer as external reference. 38

6.3.5.3 Objective Heater Calibration Press on Obj calibration icon, as Figure 38 shows, to enter the Objective Heater Calibration. Figure 44. Objective Heater calibration. To perform the calibration, refer to the following steps: 1. Stick the Temperature Sensor (green wire) at the bottom of the dish under observation; 2. Fill the dish with distilled water until the Temperature Sensor is completely immersed in water; 3. Close both this dish and the incubating chamber with their own respective lids; 4. Touch the CALIBRATE button (pointer 4 in Figure 33); 5. The warning dialog in Figure 45(a) will appear, after double checking the previous steps, click OK and the calibration will start as Figure 45 shows. Note 1. To calibrate the Objective Heater be sure that Chamber calibration has already been performed. (a) Figure 45. Objective Heater calibration. Figure 46 shows the Advanced calibration parameters for the procedure, that are accessible by pressing on Options during the calibration procedure. Figure 46. Objective Heater calibration. Advanced calibration parameters. 39

6.3.6 Logging UNO CONTROL UNIT is equipped with local memory to store the temperature and calibration data on its internal memory. These data can be then downloaded on a USB stick at user request. In order to activate the logging, follow the steps indicated below: 1. Press on Logging icon and flag Internal memory in the page that appears, see Figure 47. Tip 6. When activating the logging on the internal memory, you can access the Logging page also by pressing the activated logging icon on the Homepage, as shown in Figure 48. (a) Figure 47. Logging. (a) How to enter in the logging menu; logging in internal memory Figure 48. How to access the Logging page from the Homepage 2. Press on the icon to insert the logging time, see Figure 49. You can change this time in the range 1-60 seconds. (a) Figure 49. How to set the logging time 40

3. Choose how you want to split the data when downloaded, by pressing on Day, Week or Month, see Figure 49 (a). Tip 7. If you select Day, the data will be split in files, each one containing the data on one day. If you select Week, the data will be split in files, each one containing the data on one week. 4. Write the file name suffix to be reported in the downloaded files by pressing on the keyboard icon, see Figure 50(a). Then press on Save, see Figure 50. Tip 8. With the selections done in Figure 49 and in Figure 50, the data will be downloaded in the USB drive as shown in Figure 51. (a) Figure 50. Data Logging. How to define the suffix for the downloaded files Figure 51. Downloaded files on the pen drive Note 3. If you press on the screwdriver, see Figure 52(a) you can view the logging starting date, the available memory and the latest download, see Figure 52. Note 4. The available memory depends on the time interval you have inserted in the Logging page. The default logging time is 30 seconds. (a) Figure 52. Internal memory status. (a) Logging page settings; Internal log page settings To download the data, press on To USB or on Erase if you want to delete them, see Figure 52. 41

(a) Figure 53. Download to USB You can activate the logging also on the USB stick, by flagging on USB pen drive, see Figure 54(a). (a) Figure 54. Logging on USB drive. (a) USB drive selection; Reminder to connect the USB drive In this case, UNO advices you that a pen drive must be connected to the USB port, see Figure 54, and will store the data on the USB drive. 6.3.7 Alarms When UNO CONTROL UNIT triggers on alarm, it may activate both visual and audible alarms. To set the alarms specifications, press on the Alarms icon below: ; see Figure 55(a) then follow the indications 1. Insert the alarm time; i.e. the time for which the temperature of the Heating Devices may remain outside the allowed tolerance before the controller triggers in alarm, see 1 in Figure 55 Note 5. The alarm time range is 5-20 minutes. 2. Insert the Tolerance value, see 2 in Figure 55. This value defines the allowed tolerance form the set point. Note 6. The tolerance value range is 0.1-2.0 C 3. Insert the transient time; i.e. the maximum time that the system can take to reach a new set point, without triggering the alarm, see 3 in Figure 55. 42

(a) Figure 55. Alarms settings. (a) How to enter the Alarm setting menu; Alarm configuration Note 7. Tip 9. The transient time range is 30-240 minutes. After any change in the set point value the device enters into a transient regime. 4. Flag the tab labeled Buzzer if you want the alarm to be acoustical, additionally to being visual, see Figure 55 5. Flag the tab labeled Enabled to enable the Alarms logic, see Figure 55 6. To disable the visual and acoustical alarms, remove the flag from the tab labeled Enabled. In this case the Alarms page becomes as shown in Figure 56. Figure 56. Alarms page when the alarms are disabled Tip 10. When disabling the alarms, the CONTROLLER goes into alarm condition only when the status LED becomes red (for example is a device is accidentally disconnected) Note 8. By Pressing on Log (see Figure 57(a)), you can view all the times that the status LED of the two devices have changed color (see Figure 57). (a) Figure 57. Logging alarms page 43

Press on To USB if you want to download the alarms data to a USB drive or on Erase if you want to delete them. 6.3.8 Display To enter in the Display configuration menu, press on the icon page shown in Figure 58 will appear. 1. Options. See paragraph 6.3.8.1. 2. Brightness. See paragraph 6.3.8.2. 3. Calibration. See paragraph 6.3.8.3. 4. Date & Time. See paragraph 6.3.8.4. 5. Visual Effects. See paragraph 6.3.8.5. 6. Factory Reset. See paragraph 6.3.8.6., as shown in Figure 58(a) and the Display (a) Figure 58. Display settings page. (a) How to enter the Display menu. Display menu. 6.3.8.1 Options The Display Options menu allows t set the time frame in which the minimum and maximum temperature values are collected. To insert the time frame, press on the icon Displayed time frame bar or click the +/- icons, then press Save, see Figure 59. and slide your finger along the (a) Figure 59. Options subpage configuration. (a) Select options; Setting display options 44

Note 9. The max and minimum temperature value will be displayed in the top left box in the Homepage (see pointer 9 in Figure 17). To change the touch sound frequency press and slide your finger along the setting bar indicated with Sound frequency, then press Save, see Figure 59 Note 10. To activate a sound at each touch, flag the tab labeled Touch Sound 6.3.8.2 Brightness To modify display Brightness press the icon and scroll the bar or press the +/- icons in the page that opens, as shown in Figure 60. (a) Figure 60. Brightness display page Set the required configuration and then press on Save to obtain the right configuration of Brightness, see Figure 60. 6.3.8.3 Calibration When pressing the calibration icon, the procedure for the calibration of the touch screen will start. Figure 61. Touch screen calibration. Note 11. If the touch screen is not calibrated and it is not possible to navigate in the menu, then press and hold the power button for more than 10 seconds and the touch calibration will start. 6.3.8.4 Date & Time To set Date & Time, follow the instructions below: 1. Press the Date & Time icon, see Figure 62(a) 45

2. Set the values, as shown in Figure 62 3. Click on Save to save or Cancel to undo (a) Figure 62. Select Date&Time configuration. Tip 11. Flag the 24 hours box, if you want to use the hour format based on 24 hours. 6.3.8.5 Visual Effects To enter in the Display visual effects menu, press on the icon, see Figure 63 (a). (a) Figure 63. Display Visual Effects. (a) How to enter in Visual Effects menu; Visual Effects menu. The Top LED settings allows you to when the Top LED should be illuminated: if you select Never, the Top LED will never illuminate, if you select On Alarm, the Top LED will illuminate only when the Controller is in Alarm Status (see paragraph 6.1.1), if you select Always, the Top LED will always illuminate. Glance Mode Setting (see also paragraph 6.5) The Glance mode setting defines the color of the Top LED and of the digits displayed in Glance Mode. If you select White, the Top LED illuminates in white color (according to the criterion selected in Top LED setting) and the digits displayed in Glance Mode are white. If you select Dynamic the color of the Top LED and the color of the digits in Glance Mode follow the color coding reported in paragraph 6.1.1. Tip 12. To swap between Icon and Glance mode press on the icon. 6.3.8.6 Factory Reset To reset the display settings, press on the icon, see Figure 64(a). At this point Figure 64 will appear, asking to confirm the Factory Reset of the display. 46

(a) Figure 64. Factory reset. 6.3.9 Password With the UNO CONTROL UNIT, as well as with many Okolab s products, the user have the chance to secure his laboratory setups by using a password. The password, when enabled, will be asked, in the following procedures: Heating devices calibration procedures T sensor calibration Data download Data erase To enable the password protection, follow the steps below: 1. Press on the Password icon, see Figure 65(a) 2. Flag the tab Password enabled as shown in Figure 65 (a) Figure 65. Password. (a) Password settings menu; password subpage Note 12. The default password is okolab, and it is disabled. Okolab recommends changing this password. Note 13. If you forgot the password, use the restore password reported on the certificate attached to UNO CONTROL UNIT. 3. To change the password, press on the icon and follow the indications that will appear. 47

6.3.10 Summary The Summary Page reports technical information. To enter the Summary menu, press on the settings icon appear., as shown in Figure 66(a), then press on Summary icon. The page reported in Figure 66 will Note 14. This page is useful essentially for technical/control reasons. During a support case the engineers of Okolab s team may ask you to read some of the data reported in Summary page (a) Figure 66. Auto on/off. (a) Auto on/off menu; Auto on/off subpage. 6.3.11 Auto on/off Press on to enter the Auto on/off menu, see Figure 67(a). Once you are in the Auto on/off menu, you can choose between, see Figure 67: Auto turn on: to schedule the automatic turn on of UNO CONTROL UNIT; Auto turn on: to schedule the automatic turn on of UNO CONTROL UNIT. (a) Figure 67. Auto on/off. (a) Auto on/off menu; Auto on/off subpage. We will now describe the available options for the Auto turn on procedure only, but the description perfectly fits also to the Auto turn off procedure, whose point to point description has been omitted for the sake of simplicity. 48

Figure 68(a) shows how to access the Auto turn on subpage. As Figure 68 depicts, by default the Auto turn on/off is disabled. (a) Figure 68. Auto on/off options. It is possible to set a time when UNO CONTROL UNIT has to start Every day by pressing on its tab, or it is possible to select the specific days in the week by choosing Specific days, see Figure 69(a). Pressing Save will make your choice effective. (a) Figure 69. Auto on/off on specific days. Status On this page you can read the Status of CO2-O2-CONTROLLER; this is useful to check that the system is working properly. Most of the parameters displayed are technical/control parameters that you may be asked to access in case you have requested for Okolab support. (a) Figure 70. Status page. 49

Glance Mode Pressing on Glance Mode (see Figure 71(a)), you can change the data visualization format on the Homepage that turns into a brief information about the state of the atmosphere inside the incubator, as Figure 71 shows. (a) Figure 71. Glance Mode view. This visualization type allows you to have a quick overview of the device from far away. See paragraph 6.3.8.5 for Glance Mode settings. 50

7 MAINTENANCE To get a correct Gas controller operation over time: Use a polishing cloth or dry cloth to wipe off dust and dirt. Never use thinners, benzene, solvents on or near the devices, since these could corrode the surfaces. Verify the status of all hoses/tubing and replace those damaged. After 2 years, disconnect all polyurethane tubing, cut the last 1 cm of the tubing and reconnect them to the connectors. Before cleaning the unit, make sure it is unplugged form the power source. Water must not enter the system. 51

8 SUPPORT Support page The support page can be reached pressing on the question mark in Homepage (see Figure 72(a)), the page in Figure 72 will open. Here you can see the Italian and American Okolab contacts, moreover you can use the QR code to get to the Okolab site. (a) Figure 72. Support page. In order to send directly an email to the Okolab staff, you can use the QR codes shown in the following images: For commercial info: For technical info: Figure 73. QR code for commercial info contact. Figure 74.QR code for technical info contact. 52

Support Contacts Please, do not hesitate to contact Okolab if you need any further commercial information or technical support. Please, check Okolab web site www.oko-lab.com for news, events, new products and general FAQs. For COMMERCIAL SUPPORT: lanzaro@oko-lab.com Phone +39 081 806 2624 Fax: +39 081 876 4410 Mobile: +39 348 9680717 For TECHNICAL SUPPORT: sibillo@oko-lab.com Phone +39 081 806 3470 Mobile: +39 348 9680718 Okolab S.r.l. Via A. Olivetti, 1-80078 Pozzuoli, NA Italy Web Conference For Assistance And Training In case you requested on line support and/or web conference assistance for system assembling and experiment support, you received a web cam. Please follow these guidelines to receive optimum service. Webcam installation (instruction and Cdrom included) Latest Skype software installed (www.skype.com) Register yourself on www.skype.com to have an account (Skype_ID) Set the audio and video and test them using Skype. Contact our technical support (sibillo@oko-lab.com) by e-mail to take an appointment for the web assistance. 53

9 TECHNICAL SPECIFICATIONS UNO CONTROL UNIT Technical Specifications Operating Temperature +3 C above ambient ~.+60 C Storage Temperature -5 C ~ +60 C Operating Humidity 90% Power Consumption 24V- 5A, 120 W max Weight 0,4 Kg 54