Template talk:Oxygen sensor test

This text originated from 2 pages which were merged. The original texts can be found below:

https://wiki.oroboros.at/index.php/Oxygen_sensor_test

Figure 2. POS Quality control using the DatLab protocol (MiPNet06.03 POS-calibration-SOP)

1. Even before final equilibration, perform a stirrer test [F9], switching both stirrers automatically off and on.

2. About 20 min are required for approximate air equilibration after temperature equilibration of the incubation medium, visualized as stabilization of the Peltier power (Fig. 2; time scale is 01:10 hh:mm).

Quality control a: Upon automatic re-start of the stirrer (On), the increase of the oxygen signal should be rapid and monoexponential.

Quality control b: The raw signal (blue plot; 1 V = 1 μA at gain 1)should be close to 1 to 3 V at 25 - 37 °C at sea level up to 1,000 m altitude, in the range of pb 101 to 90 kPa (at gain setting of 2 the raw signal [V] would be two times higher).

3. Within 40 min, the oxygen signals should be stable with an O2 slope (uncorrected) close to zero (Fig. 2).

Quality control c: Signal noise should be low, reflected in a noise of the O2 slope (red plot) within ±2 (±4 is acceptable) pmol∙s⁻¹∙mL⁻¹ at a data recording interval of 2 s and 40 data points selected for calculation of the slope (Fig. 2).

4. Set a mark on the oxygen signal (R1) and click on O2 Calib. to open the DatLab O2-Calibration window.

Quality control d: The slope uncorrected should be within ±1 pmol∙s⁻¹∙mL⁻¹ averaged across the section of the experiment marked as R1 for air calibration (d). The recorded POS signal should be close to the previous calibration under identical experimental conditions. See O2-Calibration window (Fig. 2; b’).

5. Continue with a complete Instrumental O₂ background test(MiPNet14.06) or simply close the chamber and if required perform a zero oxygen calibration.

Quality control e: After closing the chamber, select plot Y2 and set mark J°1. Background slope (neg.) should be within 2.5 ± 1 pmol∙s⁻¹∙mL⁻¹.

• Flux values higher than 4.0 pmol∙s⁻¹∙mL⁻¹ indicate a biological contamination.
• Flux values lower than 1.5 pmol∙s⁻¹∙mL⁻¹:

1. Air bubbles in the Closed chamber: switch on the illumination of the O2k and inspect the O2k-Chamber through the front window. Remove any air bubbles.
2. A large volume of medium collected in the receptacle of the stopper: siphon off excess medium.
3. A larger chamber volume: check O2k-Chamber volume calibration.

Quality control f: The zero signal at mark R0 for zero calibration (not shown) should be < 2% of R1 (stable at < 5% is acceptable).

https://wiki.oroboros.at/index.php?title=Oxygen_calibration_-_DatLab&action=edit

Calibration and quality control

The POS sensor test

Stirrer test for quality control (standard 30 s) with 30 min time scale displayed with Graph Layout “02-Calibration - Background” (MiR05; 37 °C; data recording interval: 2 s; slope smooting: 40 data points).

1. About 20 min are required for approximate air equilibration after temperature equilibration of the incubation medium.
2. Even before final equilibration, perform a stirrer test [F9], switching both stirrers automatically off for 30 s.
   1. Quality control a: Upon automatic re-start of the stirrer (On), the increase of the oxygen signal should be rapid and monoexponential (see Fig. right, label a)
   2. Quality control b: The raw signal (blue plot; 1 V = 1 µA at Gain 1) should be close to 1 to 3 V at 25 to 37 °C at sea level up to 1000 m (p_b 101 to 90 kPa). At Gain setting of 2 the raw signal [V] is multiplied by 2.
3. 

   Quality control
   Within 40 min, the oxygen signals should be stable with O2 slope (uncorrected) close to zero. Quality control label c: Signal noise should be low, reflected in a noise of the O2 slope (red plot) within ±2 (±4 is acceptable) pmol/(s∙mL) at a data recording interval of 2 s and 40 data points selected for calculation of the slope.
4. Set a mark on the oxygen signal (R1) and click on O2 Calib. to open the DatLab O2 calibration. Quality control label d: The slope uncorrected should be within ±1 pmol/s∙mL) averaged across the section of the experiment marked as R1 for air calibration (d). The recorded POS signal should be close to the previous calibration under identical experimental conditions. See O2-Calibration window (see Fig. right, label label b’).
5. Continue with an instrumental O2 background test and perform a zero oxygen calibration. Quality control label label e: The zero signal at mark R0 for zero calibration should be <2 % of R1 (stable at <5 % is acceptable).