Template:Fluorescence dyes and settings

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