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Difference between revisions of "Troubleshooting: fluorescence module"

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=== Defective Fluo-Sensor or O2k or O2k-LED2 Fluo-Module===
=== Defective Fluo-Sensor or O2k or O2k-LED2 Fluo-Module===



Revision as of 14:08, 16 March 2021


                  


O2k-Open Support

Troubleshooting: fluorescence module



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MitoPedia O2k and high-resolution respirometry: O2k-Open Support 



Defective Fluo-Sensor or O2k or O2k-LED2 Fluo-Module

If the raw fluorescence signal is 0 V with and without fluorescence dyes, it might be possible that your Fluo-Sensor is defective. How can you identify the problem?
Please go through the following check-list:
1. Check that the illumination is switched off in the ´O2k control´window.
2. Check the settings for the Amperometric,Ampchannel in the ´O2k control´ window, in the Amperometric,Amp tab. Gain for Fluo-Sensor and the Fluo intensity cannot be 0.
3. The Fluo-Sensors are connected to the O2k and inserted into the O2k-Chamber in the right position (see above in the 1.3 Connect Smart Fluo-Sensor to O2k section).
4. If all the above-mentioned steps are checked and the detected signal with your Fluo-Sensor is 0 (negative or very high) in order to localise the problem, perform the following test. Add distilled water or respiration medium into the O2k-Chamber, insert the black stopper and change the settings for fluorometric measurements (see above):
  • 1. run: Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B
  • 2. run: Fluo-Sensor A in chamber B and Fluo-Sensor B in chamber A
If your Fluo-Sensor A is defective, the fluorescence signal should be 0 V with Fluo-Sensor A both in chamber A and B. In this case, the Fluo-Sensor should be shipped to us in order to repair it.
If the O2k or the O2k-LED2 Fluo-Module are defective, 0 V or negative fluorescence signal can be observed just in one chamber with two different Fluo-Sensors (see example below). In this case, the O2k or the Fluorescence- Control Unit (Series D-G) should be shipped to us in order to repair it.
Please contact our customer support to analyse your problem before shippment. Please always send us original DatLab files with the above-mentioned tests to evaluate the defective Fluo-Sensor performance.
Problem: Defective O2k or O2k-LED2 Fluo-Module
Provided by Hazirah Hassan, University Kebangsaan Malaysia, hazy_hassan@yahoo.com
Hassan Hazirah 1.test.png
Hassan Hazirah 2.test.png
Hassan Hazirah biological sample.png
Analysing these tests after changing the Fluo-Sensors between the O2k-Chambers, it is obvious that the problem is related to the O2k or O2k-LED2 Fluo-Module, since the fluorescence signal is negative and unstable in chamber B with both Fluo-Sensors. In the third figure the fluorescence signal is very unstable and it makes measurement unpossible with biological sample. Both the O2k and the LED2 Fluo-Module should be shipped to our electronic workshop.

The raw fluorescence signal [V] appears constantly at 10 V

If you see a constant raw signal at 10 V when using the O2k-Fluo Smart-Module or O2k-Fluo LED2-Module, this is probably due to high light exposure to the photodiode. Check the following parameters:
  • The chamber illumination must be switched off: Check it in the O2k control window.
  • The photodiode filters must be inserted, otherwise the light emitted from the LED will not be filtered. Click here more information on how to mount the filter cap.
  • Ensure that the Smart Fluo-Sensor is correctly inserted at the O2k frame of the chamber window.
  • Use black PEEK stoppers.

Negative fluorescence signal

Problem: The fluorescence signal of the Fluo-Sensor in chamber B is negative, while the other Fluo-Sensor has ~0 V. The same result occurs when the O2k-LED2 Fluo-Module is disconnected from the the O2k/switched off.
Provided by Anne Laure Charles, FR_Strasbourg_Zoll J,anne laure charles <anne.laure.charles@outlook.fr
Anne Laure Charles files.png
Answer: Connacting our electronic workshop, the problem might be related to a wet plug of the Fluo-Sensors. The solution is to dry it out somehow e.g. hair dryer and let it run connected to the O2k for a couple of days in dry and warm environment. If these treatments cannot solve the problem and you get still negative fluorescence values, Fluo-Sensor should be sent back to us and our electronic partner will look at it and replace it if needed.
Questions.jpg

Useful links:

» O2k-FluoRespirometer manual
» O2k-Demo experiments
» Sole source information
» Oroboros O2k