Template:Fluorescence dyes and settings
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Revision as of 13:11, 19 November 2021 by Komlodi Timea (talk | contribs)
- The light intensity of the LED (LED-intensity) and the signal amplification (Gain) can be adjusted in a wide range. The table suggests initial values, which can be optimised for specific applications.
- The settings depend on the concentration of the fluorophore, which vary between different applications. Therefore, only recommendations for specific fluorophore concentrations are given. In the Amplex® UltraRed assay the fluorophore is formed during the experiment.
- The recommendations apply to experiments at 37 °C. The Fluorescence intensity increases strongly at lower temperatures. Then the light intensity is reduced to avoid off-scale signals.
Application | Sensor | Filter set | O2k output | Light intensity (Amp polarization voltage) | Gain | Comment |
---|---|---|---|---|---|---|
AmplexTM UltraRed | Fluorescence-Sensor Green or Smart Fluo-Sensor Green | AmR | Type II | 100 - 500 | 1000 | at c(AmR) = 10 µM |
TMRM | Fluorescence-Sensor Green or Smart Fluo-Sensor Green | AmR | Type III | 100 - 500 | 1000 | at c(TMRM) = 2 µM |
Safranin | Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue | Saf | Type III | 100 - 250 | 1000 | at c(safranin) = 2 µM |
Rhodamine 123 | Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue | Saf | Type III | 100 - 250 | 1000 | at c(Rh123) = 1 µM |
Magnesium Green™ | Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue | MgG / CaG | Type I and II | 100 - 500 | 1000 | at c(Mg Green) = 2 µM |
Calcium Green™ | Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue | MgG / CaG | Type I | 300 - 500 | 1000 | at c(Ca Green) = 2 µM |
In the O2k-LED2-Module (Series D-G) polarization voltage is used, while in the O2k-Fluo Smart Module (Series H-I) fluorescence intensity. To use the same setting in the O2k-Fluo LED2-Module as in the O2k-Fluo Smart-Module use the following Excel sheet:
Download the EXCEL calculation-sheet to adjust fluorescence intensity |