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Template:Fluorescence dyes and settings

From Bioblast
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:

Download2.png Download the EXCEL calculation-sheet to adjust fluorescence intensity
Download2.png