Difference between revisions of "Troubleshooting: fluorescence module"
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{{Technical support}} | {{Technical support}} | ||
In the following page, the most common problems occurring either | In the following page, the most common problems occurring with either the [[O2k-Fluo Smart-Module]] or [[O2k-Fluo LED2-Module]] are summarized. The problem can be related to (''a'') the fluorescence settings in DatLab, (''b'') the fluorescence sensors, (''c'') the [[Fluorescence-Control Unit]] (in the case of the [[O2k-Fluo LED2-Module]]), or (''d'') the O2k. | ||
__TOC__ | __TOC__ | ||
== The raw fluorescence signal is 0 V | == Zero fluorescence signal == | ||
::: '''Problem:''' The raw fluorescence signal is 0 V with and without the fluorescence dye. | |||
:::'''Solution:''' To identify cause of the problem, please go through the following check-list: | |||
:::: 1. Check that the Fluo-Sensors are connected to the O2k and inserted into the O2k-chamber in the right position. See: [[O2k-Fluo_Smart-Module#Connect_Fluorescence-Sensor_to_O2k |O2k-Fluo_Smart-Module#Connect_Fluorescence-Sensor_to_O2k]]. | |||
::: | :::: 2. Check that the [[Fluorescence-Control Unit]] is properly connected to the O2k of series D-G. | ||
:::: | :::: 3. In DatLab check that: | ||
::::: a) the [[Illumination on/off|illumination]] is switched off in the ´[[O2k control]]´window. | |||
::::: b) the gain for Fluo-Sensor and the Fluo intensity are set to a number greater than 0 in the 'Amperometric,Amp' tab of the ´[[O2k control]]´ window. | |||
:::: 4. If all the above-mentioned settings and set up are correct and the detected signal with the Fluo-Sensor is still 0, perform the following test to localize the problem: | |||
:::: 4. If all the above-mentioned | ::::::Add distilled water or respiration medium into the O2k-chamber (without sample and fluorescence dye), insert the black stopper and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Serie H or higher) or Polarisation voltage (Series D-G)=500. | ||
::::::* | ::::::* Let it run for 5 min with Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B. | ||
::::::* | ::::::* Swap the Fluo-sensors and set event “swap sensor”, write in the comment box which sensor is in which chamber. | ||
:::::: If | ::::::* Let it run for 5 min. | ||
:::::: If the O2k or the O2k-LED2 Fluo-Module | :::::: If the problem is with the Fluo-Sensor, the fluorescence signal will be 0 V with the faulty Fluo-Sensor in both chambers. In this case, the Fluo-Sensor should be shipped to us for repairs. | ||
::::: Please contact our [https:// | :::::: If the problem is with the O2k or the O2k-LED2 Fluo-Module, 0 V fluorescence signal can be observed just in one chamber with two different Fluo-Sensors. In this case, the O2k or the Fluorescence-Control Unit (Series D-G) should be shipped to us for repairs. In some cases, the defect of the Fluorescence-Control Unit (Series D-G) leads to 0 V in both chambers. | ||
:::: '''Example: | ::::: Please contact our [https://www.oroboros.at/index.php/o2k-technical-support/ O2k-technical Support] to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance. | ||
== Noisy fluorescence signal == | |||
::: '''Problem:''' The raw fluorescence signal is noisy to such an extent that the evaluation of the results is not possible. | |||
:::'''Solution:''' To localize the problem, perform the following test: | |||
::::* Add distilled water or respiration medium into the O2k-chamber (without sample and fluorescence dye), insert the black stopper and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Serie H or higher) or Polarisation voltage (Series D-G)=500. | |||
::::::* Let it run for 5 min with Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B. | |||
::::::* Swap the Fluo-sensors and set event “swap sensor”, write in the comment box which sensor is in which chamber. | |||
::::::* Let it run for 5 min. | |||
:::::: If the problem is with the Fluo-Sensor, the fluorescence signal will be noisy with the faulty Fluo-Sensor in both chambers. In this case, the Fluo-Sensor should be shipped to us for repairs. | |||
:::::: If the problem is with the O2k or the O2k-LED2 Fluo-Module, the fluorescence signal is noisy 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 for repairs. If the noise occurs in both chambers, the Fluo-Sensors and the O2k (and Fluorescence-Control Unit - series D-G) should be shipped to us. | |||
::::: Please contact our [https://www.oroboros.at/index.php/o2k-technical-support/ O2k-technical Support] to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance. | |||
:::: '''Example: Problem with O2k or O2k-LED2 Fluo-Module''' | |||
:::::Provided by Hazirah Hassan, University Kebangsaan Malaysia, hazy_hassan@yahoo.com | :::::Provided by Hazirah Hassan, University Kebangsaan Malaysia, hazy_hassan@yahoo.com | ||
::::[[File:Hassan Hazirah 1.test.png|700px]] | ::::[[File:Hassan Hazirah 1.test.png|700px]] | ||
::::[[File:Hassan Hazirah 2.test.png|700px]] | ::::[[File:Hassan Hazirah 2.test.png|700px]] | ||
::::[[File:Hassan Hazirah biological sample.png|700px]] | ::::[[File:Hassan Hazirah biological sample.png|700px]] | ||
:::: After changing the Fluo-Sensors between the O2k-chambers, 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 in chamber B, which makes the experiment hard to interpret. Both the O2k and the LED2 Fluo-Module should be shipped to our electronic workshop. | :::: After changing the Fluo-Sensors between the O2k-chambers, 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 in chamber B, which makes the experiment hard to interpret. Both the O2k and the LED2 Fluo-Module should be shipped to our electronic workshop. | ||
== Negative fluorescence signal== | |||
::::'''Problem:''' The fluorescence signal of one Fluo-Sensor is negative. | |||
:::: '''Solution:''' To localize the problem, perform the following test: | |||
::::* Add distilled water or respiration medium (without sample and fluorescence dye) into the O2k-chamber, insert the black stopper, and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Series H or higher) or Polarisation voltage (Series D-G) = 500. | |||
::::::* Let it run for 5 min with Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B. | |||
::::::* Swap the Fluo-sensors and set event “swap sensor”, write in the comment box which sensor is in which chamber. | |||
::::::* Let it run for 5 min. | |||
:::::: The problem is related to one of the Fluo-Sensors, if you observe a negative signal in both chambers after swapping them. The negative signal might be related to a wet plug of the Fluo-Sensor. The solution is to dry it out with ''e.g.'' a hair dryer and let it run connected to the O2k (25 °C, illumination off, distilled water in the O2k-chamber, gain for Fluo-Sensor 1000, fluorescence intensity=500) for a couple of days in a dry and warm environment. If this treatment does not solve the problem and you get still negative fluorescence values, the Fluo-Sensor should be sent back to us for further investigation. | |||
::: Please contact our [[O2k-technical support]] to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance. | |||
:::: '''Example:''' Provided by Anne Laure Charles, FR_Strasbourg_Zoll J,anne laure charles <anne.laure.charles@outlook.fr | |||
::::[[File:Anne Laure Charles files.png|700px]] | |||
:::: The fluorescence signal was negative in both chambers after swapping the Fluo-Sensors, but the signal changed after swapping the sensors, which means that the problem is located to the Fluo-Sensors. In this case, both Fluo-Sensors should be shipped to the electronic workshop. | |||
== | == Fluorescence signal is constantly at 10 V== | ||
::: '''Problem:''' The fluorescence signal is constantly at 10 V when using the [[O2k-Fluo Smart-Module]] or [[MiPNet17.05 O2k-Fluo LED2-Module | O2k-Fluo LED2-Module]]. | ::: '''Problem:''' The fluorescence signal is constantly at the 10 V upper limit when using the [[O2k-Fluo Smart-Module]] or [[MiPNet17.05 O2k-Fluo LED2-Module | O2k-Fluo LED2-Module]]. | ||
::: '''Solution:''' This is probably due to high light exposure to the photodiode. Check the following parameters: | ::: '''Solution:''' This is probably due to high light exposure to the photodiode. Check the following parameters: | ||
:::: | ::::1. The chamber illumination must be switched off. Check it in the [[O2k control]] window. | ||
:::: | ::::2. The photodiode filters must be inserted, otherwise the light emitted from the LED will not be filtered. Information on how to mount the filter cap- » [[Filter-Cap#Mounting_a_Filter-Cap|here]]. | ||
:::: | ::::3. [[O2k-Fluo_Smart-Module#Stoppers|Black PEEK stoppers]] must be used. | ||
:::: | ::::4. The fluorescence intensity cannot be set too high. Check in the [[Amperometric,Amp]] tab of the ´[[O2k control]]´ window. | ||
::: If all the above-mentioned settings and set up are correct and the detected signal is still at 10 V (out of scale), the O2k (and Fluorescence-Control Unit - series D-G) or/and the Fluo-Sensors should be shipped to us. | |||
::: Please contact our [https://www.oroboros.at/index.php/o2k-technical-support/ O2k-technical Support] to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance. | |||
== | == High fluorescence signal == | ||
::::'''Problem:''' The fluorescence signal is suddenly higher in one or both chambers even before adding dye or sample. | |||
::::'''Problem:''' The fluorescence signal of the Fluo- | ::::'''Solution:''' The problem might be related to the position of the filters inserted onto the Fluo-Sensors. First, check whether the filters are in the correct position and have not shifted. If this does not solve the problem, perform the following test to localize the problem: | ||
:::: | ::::::Add distilled water or respiration medium into the O2k-chamber (without sample and fluorescence dye), insert the black stopper and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Serie H or higher) or Polarisation voltage (Series D-G)=500. | ||
:::: | ::::::* Let it run for 5 min with Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B. | ||
:::: | ::::::* Swap the Fluo-sensors and set event “swap sensor”, write in the comment box which sensor is in which chamber. | ||
::::::* Let it run for 5 min. | |||
:::::: If the problem is with one of the Fluo-Sensors, the fluorescence signal will be high with the faulty Fluo-Sensor in both chambers. In this case, the Fluo-Sensor should be shipped to us for repairs. | |||
:::::: If the problem is with the O2k or the O2k-LED2 Fluo-Module, the fluorescence signal is high in one chamber with two different Fluo-Sensors. In this case, the O2k or the Fluorescence-Control Unit (Series D-G) should be shipped to us for repairs. If the signal is high in both chambers, the Fluo-Sensors and the O2k (and Fluorescence-Control Unit - series D-G) should be shipped to us. | |||
::::: Please contact our [https://www.oroboros.at/index.php/o2k-technical-support/ O2k-technical Support] to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance. | |||
:::: For more information on how to assemble the Fluo-Sensors and how to change the filters, see: | |||
::::» [https://www.youtube.com/watch?v=nlVLXcbBRqs Smart Fluo-Sensor assembly] | |||
::::» [https://www.youtube.com/watch?v=eUukvgAi1yY Exchanging a filter set of the Fluo-Sensors] | |||
[[File:Questions.jpg|left|40px]] | [[File:Questions.jpg|left|40px]] | ||
== Useful links == | == Useful links == | ||
Line 51: | Line 84: | ||
::::» [[Calcium Green]] | ::::» [[Calcium Green]] | ||
::::» [[MiPNet22.11 O2k-FluoRespirometer manual| O2k-FluoRespirometer manual]] | ::::» [[MiPNet22.11 O2k-FluoRespirometer manual| O2k-FluoRespirometer manual]] | ||
::::» [[O2k-Fluo LED2-Module]] | |||
::::» [[O2k-Demo_experiments |O2k-Demo experiments]] | ::::» [[O2k-Demo_experiments |O2k-Demo experiments]] | ||
::::» [[MiPNet06.05 Specifications | Sole source information]] | ::::» [[MiPNet06.05 Specifications | Sole source information]] | ||
::::» [[Oroboros O2k]] | ::::» [[Oroboros O2k]] | ||
::::» [https://www.youtube.com/watch?v=nlVLXcbBRqs Smart Fluo-Sensor assembly] | |||
::::» [https://www.youtube.com/watch?v=eUukvgAi1yY Exchanging a filter set of the Fluo-Sensors] |
Revision as of 09:32, 31 May 2021
Troubleshooting: fluorescence module |
MitoPedia O2k and high-resolution respirometry:
O2k-Open Support
In the following page, the most common problems occurring with either the O2k-Fluo Smart-Module or O2k-Fluo LED2-Module are summarized. The problem can be related to (a) the fluorescence settings in DatLab, (b) the fluorescence sensors, (c) the Fluorescence-Control Unit (in the case of the O2k-Fluo LED2-Module), or (d) the O2k.
Zero fluorescence signal
- Problem: The raw fluorescence signal is 0 V with and without the fluorescence dye.
- Solution: To identify cause of the problem, please go through the following check-list:
- 1. Check that the Fluo-Sensors are connected to the O2k and inserted into the O2k-chamber in the right position. See: O2k-Fluo_Smart-Module#Connect_Fluorescence-Sensor_to_O2k.
- 2. Check that the Fluorescence-Control Unit is properly connected to the O2k of series D-G.
- 3. In DatLab check that:
- a) the illumination is switched off in the ´O2k control´window.
- b) the gain for Fluo-Sensor and the Fluo intensity are set to a number greater than 0 in the 'Amperometric,Amp' tab of the ´O2k control´ window.
- 4. If all the above-mentioned settings and set up are correct and the detected signal with the Fluo-Sensor is still 0, perform the following test to localize the problem:
- Add distilled water or respiration medium into the O2k-chamber (without sample and fluorescence dye), insert the black stopper and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Serie H or higher) or Polarisation voltage (Series D-G)=500.
- Let it run for 5 min with Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B.
- Swap the Fluo-sensors and set event “swap sensor”, write in the comment box which sensor is in which chamber.
- Let it run for 5 min.
- If the problem is with the Fluo-Sensor, the fluorescence signal will be 0 V with the faulty Fluo-Sensor in both chambers. In this case, the Fluo-Sensor should be shipped to us for repairs.
- If the problem is with the O2k or the O2k-LED2 Fluo-Module, 0 V fluorescence signal can be observed just in one chamber with two different Fluo-Sensors. In this case, the O2k or the Fluorescence-Control Unit (Series D-G) should be shipped to us for repairs. In some cases, the defect of the Fluorescence-Control Unit (Series D-G) leads to 0 V in both chambers.
- Add distilled water or respiration medium into the O2k-chamber (without sample and fluorescence dye), insert the black stopper and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Serie H or higher) or Polarisation voltage (Series D-G)=500.
- Please contact our O2k-technical Support to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance.
- 3. In DatLab check that:
Noisy fluorescence signal
- Problem: The raw fluorescence signal is noisy to such an extent that the evaluation of the results is not possible.
- Solution: To localize the problem, perform the following test:
- Add distilled water or respiration medium into the O2k-chamber (without sample and fluorescence dye), insert the black stopper and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Serie H or higher) or Polarisation voltage (Series D-G)=500.
- Let it run for 5 min with Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B.
- Swap the Fluo-sensors and set event “swap sensor”, write in the comment box which sensor is in which chamber.
- Let it run for 5 min.
- If the problem is with the Fluo-Sensor, the fluorescence signal will be noisy with the faulty Fluo-Sensor in both chambers. In this case, the Fluo-Sensor should be shipped to us for repairs.
- If the problem is with the O2k or the O2k-LED2 Fluo-Module, the fluorescence signal is noisy 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 for repairs. If the noise occurs in both chambers, the Fluo-Sensors and the O2k (and Fluorescence-Control Unit - series D-G) should be shipped to us.
- Please contact our O2k-technical Support to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance.
- Example: Problem with O2k or O2k-LED2 Fluo-Module
- Provided by Hazirah Hassan, University Kebangsaan Malaysia, hazy_hassan@yahoo.com
- After changing the Fluo-Sensors between the O2k-chambers, 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 in chamber B, which makes the experiment hard to interpret. Both the O2k and the LED2 Fluo-Module should be shipped to our electronic workshop.
Negative fluorescence signal
- Problem: The fluorescence signal of one Fluo-Sensor is negative.
- Solution: To localize the problem, perform the following test:
- Add distilled water or respiration medium (without sample and fluorescence dye) into the O2k-chamber, insert the black stopper, and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Series H or higher) or Polarisation voltage (Series D-G) = 500.
- Let it run for 5 min with Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B.
- Swap the Fluo-sensors and set event “swap sensor”, write in the comment box which sensor is in which chamber.
- Let it run for 5 min.
- The problem is related to one of the Fluo-Sensors, if you observe a negative signal in both chambers after swapping them. The negative signal might be related to a wet plug of the Fluo-Sensor. The solution is to dry it out with e.g. a hair dryer and let it run connected to the O2k (25 °C, illumination off, distilled water in the O2k-chamber, gain for Fluo-Sensor 1000, fluorescence intensity=500) for a couple of days in a dry and warm environment. If this treatment does not solve the problem and you get still negative fluorescence values, the Fluo-Sensor should be sent back to us for further investigation.
- Please contact our O2k-technical support to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance.
- Example: Provided by Anne Laure Charles, FR_Strasbourg_Zoll J,anne laure charles <anne.laure.charles@outlook.fr
- The fluorescence signal was negative in both chambers after swapping the Fluo-Sensors, but the signal changed after swapping the sensors, which means that the problem is located to the Fluo-Sensors. In this case, both Fluo-Sensors should be shipped to the electronic workshop.
Fluorescence signal is constantly at 10 V
- Problem: The fluorescence signal is constantly at the 10 V upper limit when using the O2k-Fluo Smart-Module or O2k-Fluo LED2-Module.
- Solution: This is probably due to high light exposure to the photodiode. Check the following parameters:
- 1. The chamber illumination must be switched off. Check it in the O2k control window.
- 2. The photodiode filters must be inserted, otherwise the light emitted from the LED will not be filtered. Information on how to mount the filter cap- » here.
- 3. Black PEEK stoppers must be used.
- 4. The fluorescence intensity cannot be set too high. Check in the Amperometric,Amp tab of the ´O2k control´ window.
- If all the above-mentioned settings and set up are correct and the detected signal is still at 10 V (out of scale), the O2k (and Fluorescence-Control Unit - series D-G) or/and the Fluo-Sensors should be shipped to us.
- Please contact our O2k-technical Support to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance.
High fluorescence signal
- Problem: The fluorescence signal is suddenly higher in one or both chambers even before adding dye or sample.
- Solution: The problem might be related to the position of the filters inserted onto the Fluo-Sensors. First, check whether the filters are in the correct position and have not shifted. If this does not solve the problem, perform the following test to localize the problem:
- Add distilled water or respiration medium into the O2k-chamber (without sample and fluorescence dye), insert the black stopper and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Serie H or higher) or Polarisation voltage (Series D-G)=500.
- Let it run for 5 min with Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B.
- Swap the Fluo-sensors and set event “swap sensor”, write in the comment box which sensor is in which chamber.
- Let it run for 5 min.
- If the problem is with one of the Fluo-Sensors, the fluorescence signal will be high with the faulty Fluo-Sensor in both chambers. In this case, the Fluo-Sensor should be shipped to us for repairs.
- If the problem is with the O2k or the O2k-LED2 Fluo-Module, the fluorescence signal is high in one chamber with two different Fluo-Sensors. In this case, the O2k or the Fluorescence-Control Unit (Series D-G) should be shipped to us for repairs. If the signal is high in both chambers, the Fluo-Sensors and the O2k (and Fluorescence-Control Unit - series D-G) should be shipped to us.
- Add distilled water or respiration medium into the O2k-chamber (without sample and fluorescence dye), insert the black stopper and change the settings for fluorometric measurements: Gain=1000; Fluo intensity (Serie H or higher) or Polarisation voltage (Series D-G)=500.
- Please contact our O2k-technical Support to analyze your problem before shipment. Be sure to always send us original DatLab files with the above-mentioned tests to evaluate Fluo-Sensor performance.
- For more information on how to assemble the Fluo-Sensors and how to change the filters, see:
- » Smart Fluo-Sensor assembly
- » Exchanging a filter set of the Fluo-Sensors