User Manual for UNISENSE models including: 2023.05 O2 Calibration Kit, 2023.05, O2 Calibration Kit, Calibration Kit, Kit
Manuals - Unisense
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DocumentDocumentO2 sensor calibration kit For electrochemical and optical oxygen sensors Manual Contents 1 WARRANTY AND LIABILITY ......................................................................................................................3 1.1 NOTICE TO PURCHASER ............................................................................................................................... 3 1.2 WARNING ................................................................................................................................................ 3 1.3 WARRANTY AND LIABILITY ........................................................................................................................... 3 2 SUPPORT, ORDERING, AND CONTACT INFORMATION .............................................................................3 3 CONTENT OF THE CALIBRATION KIT.........................................................................................................3 4 PRINCIPLE OF CALIBRATION ....................................................................................................................4 5 CALIBRATION PROCEDURE ......................................................................................................................4 5.1 IMPORTANT NOTES FOR CALIBRATING ALL UNISENSE O2 SENSORS ........................................................................ 4 5.2 CALIBRATING MOST ELECTROCHEMICAL O2 SENSORS ......................................................................................... 5 5.2.1 Obtaining the high calibration point................................................................................................. 5 5.2.1.1 Using the Unisense Cal300 Calibration Chamber.....................................................................................5 5.2.1.2 Using the calibration cap ......................................................................................................................... 6 5.2.2 Obtaining the zero-calibration point................................................................................................. 6 5.3 CALIBRATING ELECTROCHEMICAL AND OPTICAL O2 SENSORS IN FLOW CELLS ........................................................... 6 5.3.1 Create a calibration setup................................................................................................................. 6 5.4 CALIBRATING ELECTROCHEMICAL AND OPTICAL O2 SENSORS FOR THE MICRORESPIRATION SYSTEM ............................. 7 5.4.1 Obtaining the high calibration point................................................................................................. 7 5.4.1.1 Using the Unisense Cal300 Calibration Chamber.....................................................................................7 5.4.1.2 Using a Microrespiration chamber...........................................................................................................7 5.4.2 Obtaining the zero-calibration point................................................................................................. 7 5.5 CALIBRATING O2 MICROOPTODES - RETRACTABLE NEEDLE DESIGN ...................................................................... 8 5.5.1 Obtaining the high calibration point................................................................................................. 8 5.5.1.1 Using the Unisense Cal300 Calibration Chamber.....................................................................................8 5.5.1.2 Using the calibration cap ......................................................................................................................... 8 5.5.2 Obtaining the zero-calibration point................................................................................................. 9 5.6 CALIBRATING O2 MINIOPTODES - OPTO-3000................................................................................................ 9 5.6.1 Obtaining the high calibration point................................................................................................. 9 5.6.2 Obtaining the zero-calibration point................................................................................................. 9 6 SPECIFICATIONS ......................................................................................................................................9 7 EQUILIBRIUM O2 CONCENTRATION AT KNOW TEMPERATURE AND SALINITY .......................................10 O2 CALIBRATION KIT USER MANUAL Copyright © Unisense A/S Version: May 2023 2 1 Warranty and liability 1.1 Notice to Purchaser This product is for research use only. Not for use in human diagnostic or therapeutic procedures. 1.2 Warning Microsensors have very pointed tips and must be handled with care to avoid personal injury and only by trained personnel. Unisense A/S recommends users to attend instruction courses to ensure proper use of the products. 1.3 Warranty and Liability The Oxygen Calibration Kit is guaranteed to give a calibration solution with zero oxygen until expiry as indicated on the package label. The warranty does not include replacement necessitated by accident, neglect, misuse, unauthorized repair, or modification of the product. In no event will Unisense A/S be liable for any direct, indirect, consequential or incidental damages, including lost profits, or for any claim by any third party, arising out of the use, the results of use, or the inability to use this product. 2 Support, ordering, and contact information If you wish to order additional products or if you encounter any problems and need scientific or technical assistance, please do not hesitate to contact our sales and support team. We will respond to your inquiry within one working day. E-mail: sales@unisense.com Unisense A/S Langdyssen 5 DK-8200 Aarhus N, Denmark Tel: +45 8944 9500 Fax: +45 8944 9549 Further documentation and support are available at our website: www.unisense.com. 3 Content of the calibration kit Item Exetainer with zero O2 solution Calibration cap with O-ring and 3 cm Viton tubing 10 ml syringe 80 x 2.1 mm needle (green) Number 10 1 1 1 3 A B C D E Figure 1: Calibration kit contents: A: Calibration kit box with Exetainers, B: 80 x 2.1 mm needle (green), C: 10 ml syringes, D: Calibration Cap with tubing, E: O-ring. 4 Principle of calibration Unisense electrochemical O2 sensors respond linearly to O2 concentrations within their linear range (see specifications for your sensor at https://www.unisense.com/O2). Likewise, the processed signal of Unisense optical oxygen sensors (optodes) respond linearly to O2 concentrations (see https://www.unisense.com/MicroOptode). Therefore, a two-point calibration is sufficient for both types of sensors, one point is the signal for zero O2 and the other point is the signal for a known O2 concentration. This calibration kit contains Exetainers with a zero O2 solution. A solution with a known O2 concentration can be made by bubbling water, at a known temperature and salinity, with atmospheric air. The O2 concentration at equilibrium can be looked up in a table (Table 1) or by using the built in O2 solubility function in SensorTrace Suite. This is available through the O2 table button in the SensorTrace calibration routine and through the main menu in the software: Tools -> O2 calculator (See the SensorTrace Suite manual for details: https://www.unisense.com/manuals/). 5 Calibration procedure 5.1 Important notes for calibrating all Unisense O2 sensors · The temperature of the two calibration solutions must be the same. · The pre-polarization of electrochemical O2 sensors must have been completed before doing the calibration. See the O2 sensor manual for details: https://www.unisense.com/manuals/ · It is recommended to use the Unisense Calibration Chamber for obtaining the high calibration point (e.g., air saturation). · If doing the high calibration point with the calibration cap, make sure to do this before the zero calibration to avoid carry over from the zero-calibration solution. Wash the calibration cap and the protection tube very well with water after exposing it to the zero-solution. 4 Figure 2:Oxygen microsensor with the calibration cap mounted. Calibration solution is injected with the 10 ml syringe. 5.2 Calibrating most electrochemical O2 sensors (All sensors except those in Flow Cells and for the MicroRespiration System - see 5.3 & 5.4) For calibrating an electrochemical and optical O2 sensors, a low and a high calibration point are needed. 5.2.1 Obtaining the high calibration point 5.2.1.1 Using the Unisense Cal300 Calibration Chamber 1. Place the sensor with the protection tube in air saturated water at a known temperature (see the O2 microsensor manual (https://www.unisense.com/manuals/) 2. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/) 5 5.2.1.2 Using the calibration cap 1. Mount the calibration cap on the protection tube with the O2 sensor. Make sure that the Oring is in place at the bottom of the calibration cap creating a seal between this and the protection tube. 2. Fill the 10 ml syringe with air saturated water. 3. Inject this water into the calibration cap until the sensor tip is immersed at least 2-3 cm. 4. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/) 5.2.2 Obtaining the zero-calibration point 1. Mount the calibration cap, if not already mounted, as described above in 5.2.1.2 2. Open the Exetainer with the zero solution 3. Aspirate ca. 10 ml of the zero solution with the syringe and needle. 4. Keep the syringe vertical and avoid mixing of the zero solution with the air bubble inside. 5. Remove the needle and attach the 10 ml syringe to the calibration cap tubing. 6. Inject the calibration solution slowly until the sensor tip is immersed at least 2-3 cm. 7. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/) 8. Remove the zero solution with the syringe. 9. Wash the calibration cap and protection tube carefully, removing all of the zero solution. 5.3 Calibrating electrochemical and optical O2 sensors in flow cells Sensors with flow cells cannot be calibrated using the Calibration Cap. Instead, it is recommended to follow the procedure outlined below and to pay attention to the general information in section 5.1. 5.3.1 Create a calibration setup In order to calibrate a sensor in a flow cell, the sensor tip must be exposed to the calibration liquid. The optimal way to do this depends on the actual setup, however, it is recommended to make a setup that allows calibration of the sensor without removing it from the flow cell and from the setup. Generally, this could be a Luer connector connected to the flow cell, directly or via tubing, that allows injection of the calibration liquid into the flow cell. A three-way valve on either side of the flow cell will allow easy injection of the calibration liquid with the sensor and flow cell in place. Connection of the syringe with calibration liquid to the flow cells: · PEEK flow cells: The syringe may be attached directly to the flow cell via a Luer adapter that is mounted directly in the flow cell (Figure 3, left) · Glass and Swagelok stainless steel flow cells: The syringe may be attached via rubber tubing. The syringe may be connected directly to the rubber tubing or via a barbed Luer adaptor (Figure 3, right) Figure 3: Left: Luer adaptor for direct mounting in the flow cell (e.g., IDEX P-624). Right: Barbed Luer adaptor for tube connection 6 5.4 Calibrating electrochemical and optical O2 sensors for the Microrespiration system Sensors of the Microrespiration type (OX-MR) cannot be calibrated using the calibration cap. Instead, it is recommended to use the MR zero calibration chamber as described below. Figure 4: Oxygen sensor in the Microrespiration guide. 5.4.1 Obtaining the high calibration point 5.4.1.1 Using the Unisense Cal300 Calibration Chamber 1. Place the sensor in the Cal300 Calibration Chamber containing air saturated water at a known temperature (see the O2 microsensor manual (https://www.unisense.com/manuals/). · The O2 sensor must be mounted in the blue Microrespiration guide, and the tip must be retracted. · Temperature and salinity of the water must be the same as where the measurements are done. 2. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/) 5.4.1.2 Using a Microrespiration chamber 1. Equilibrate water with atmospheric air by bubbling. The temperature and salinity of this must be the same as where the measurements are done. 2. Transfer the air saturated water to a MicroRespiration chamber and put the lid on the chamber. 3. Place the MicroRespiration chamber in the stirrer rack. It is recommended to place this in a water bath to keep control over the temperature. 4. Place the sensor in the stirrer rack with the plastic tip in the opening of the Microrespiration chamber lid and insert the O2 sensor tip into the MicroRespiration chamber. 5. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/). 5.4.2 1. 2. 3. 4. 5. 6. Obtaining the zero-calibration point Fill the silicone tubing, attached to the zero-calibration chamber lid, with water. Make sure all bubbles are removed. Fill the 4 ml zero calibration chamber with the zero O2 solution. Mount the lid on the zero-calibration chamber taking care to avoid bubbles (see note A below). Place the zero-calibration chamber at the single position, closest to the cable in the stirrer rack. Place the sensor in the stirrer rack with the plastic tip in the opening of the Microrespiration chamber lid and insert the O2 sensor tip into the MicroRespiration chamber. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/). 7 NOTE: The water filled silicone tubing is now placed in the zero O2 solution and O2 from inside the silicone tubing will diffuse into the ascorbate solution and be consumed. It is recommended to leave the zerocalibration chamber over night for complete removal of O2 from the water inside the silicone tubing. Figure 5: Zero calibration chamber (MR-Ch 0-calibration). The zero solution is outside the silicone tubing and pure water is inside. The oxygen in the water inside the tubing diffuses through the silicone tubing into the zero solution where it is consumed. 5.5 Calibrating O2 MicroOptodes - Retractable Needle design NOTE: Before calibrating optodes, the optical fibre must be extended maximally out of the needle. This is done by pushing the inner steel cylinder into the black shaft. Figure 6: Optical oxygen sensor, Optode, in the Retractable Needle design. 5.5.1 Obtaining the high calibration point 5.5.1.1 Using the Unisense Cal300 Calibration Chamber 1. Place the sensor with the protection tube in air saturated water at a known temperature (see the MicroOptode - Opto Series - Manual (https://www.unisense.com/manuals/) 2. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/) 5.5.1.2 Using the calibration cap 1. Mount the calibration cap on the protection tube with the O2 sensor. Make sure that the Oring is in place at the bottom of the calibration cap creating a seal between this and the protection tube. 2. Fill the 10 ml syringe with air saturated water. 3. Inject this water into the calibration cap until the sensor tip is immersed. 4. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/). 8 5.5.2 Obtaining the zero-calibration point 1. Mount the calibration cap, if not already mounted, as described above in 5.5.1.2 2. Open the Exetainer with the zero solution 3. Aspirate ca. 10 ml of the zero solution with the syringe and needle. 4. Keep the syringe vertical and avoid mixing of the zero solution with the air bubble inside. 5. Remove the needle and attach the 10 ml syringe to the calibration cap tubing. 6. Inject the calibration solution slowly until the sensor fibre tip is well immersed and stop before the solution reaches the needle. If the ascorbate solution gets into the needle, it is difficult to wash out. 7. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/) 8. Remove the zero solution with the syringe. 9. Wash the calibration cap, sensor tip and protection tube carefully, removing all of the zero solution. 5.6 Calibrating O2 MiniOptodes - Opto-3000 5.6.1 Obtaining the high calibration point 1. Place the sensor in air saturated water at a known temperature (see the MicroOptode - Opto Series - Manual (https://www.unisense.com/manuals/) 2. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/) 5.6.2 Obtaining the zero-calibration point 1. Open the Exetainer with the zero solution. 2. Insert the Opto-3000 sensor into the zero solution at least 5 mm. Use a clamp or other holder to keep the sensor steady. Take care that air bubbles are not trapped at the front of the sensor. 3. Allow the sensor to respond and stabilize and record the calibration value in SensorTrace (see the SensorTrace manual for details: https://www.unisense.com/manuals/) 4. Wash the sensor carefully, removing all of the zero solution. 6 Specifications · Content of the zero-calibration solution: · Volume of the zero-calibration solution: · Lifetime of the calibration kit: Slightly alkaline ascorbate 12 ml See label on the calibration kit box Figure 7: Optical oxygen sensor, Optode, in the Opto-3000 design. 9 7 Equilibrium O2 concentration at know temperature and salinity Detailed tables are available at: http://www.unisense.com/technical_information/ At 20°C and 1 atm.: 1 µmol O2/l = 0.032 mg O2/l = 0.024 ml O2 Table 1: Concentrations of oxygen (µmol O2/litre) in water equilibrated with moist atmospheric air at a partial pressure of 0.21 atm. as a function of temperature and salinity. °C 0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0 16.0 18.0 20.0 22.0 24.0 26.0 28.0 30.0 32.0 34.0 36.0 38.0 40.0 42.0 0.0 456.6 450.4 444.2 438.1 432.1 426.1 420.3 414.5 408.8 403.2 397.7 392.2 386.8 381.5 376.2 371.0 365.9 3609 355.9 351.0 346.2 341.4 5.0 398.9 393.6 388.5 383.3 378.3 373.3 368.4 363.5 358.7 354.0 349.3 344.7 340.2 335.7 331.2 326.9 322.5 3183 314.1 309.9 305.8 301.8 10.0 352.6 348.1 343.7 339.4 335.1 330.8 326.7 322.5 318.4 314.4 310.4 306.5 302.6 298.7 294.9 291.2 287.5 2839 280.3 276.7 273.2 269.4 15.0 314.9 311.1 307.3 303.6 299.9 296.2 292.6 289.1 285.5 282.1 278.6 275.2 271.9 268.5 265.3 262.0 258.8 2557 252.5 249.5 246.4 243.4 20.0 283.9 280.6 277.3 274.0 270.8 267.6 264.5 261.4 258.3 255.3 252.3 249.3 246.4 243.5 240.6 237.8 235.0 2322 229.5 226.8 224.1 221.5 25.0 257.9 255.0 252.1 249.3 246.5 243.7 240.9 238.2 235.5 232.8 230.2 227.6 225.0 222.5 219.9 217.4 215.0 2125 210.1 207.7 205.4 203.1 30.0 235.9 233.3 230.8 228.3 225.8 223.3 220.9 218.5 216.1 213.7 211.4 209.1 206.8 204.5 202.3 200.1 197.9 1957 193.6 191.4 189.3 187.3 35.0 217.0 214.7 212.4 210.2 207.9 205.7 203.6 201.4 199.3 197.2 195.1 193.0 191.0 189.0 187.0 185.0 183.0 1811 179.2 177.3 175.4 173.6 40.0 200.4 198.3 196.3 194.3 192.3 190.3 188.4 186.5 184.6 182.7 180.8 179.0 177.1 175.3 173.5 171.7 170.0 1682 166.5 164.8 163.1 161.5 Unisense, Denmark www.unisense.com - sales@unisense.com 10
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