Template:Fluorescence dyes and settings: Difference between revisions
From Bioblast
mNo edit summary |
No edit summary |
||
| (3 intermediate revisions by 2 users not shown) | |||
| Line 1: | Line 1: | ||
{| class="wikitable" | :::: The light intensity of the LED ([[Fluorescence-Control_Unit#Control_of_LED-intensity|LED-intensity]]) and the signal amplification ([[Fluorescence-Control_Unit#Gain|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. | |||
{| class="wikitable" | |||
|- | |- | ||
! Application !!Sensor!! Filter set !! O2k output !! Light intensity (Amp polarization voltage) !! Gain !! Comment | ! Application !!Sensor!! Filter set !! O2k output !! Light intensity (Amp polarization voltage) !! Gain !! Comment | ||
| Line 5: | Line 8: | ||
| [[Amplex UltraRed|Amplex<sup>TM</sup> UltraRed]]||[[Fluorescence-Sensor Green]] or [[Smart Fluo-Sensor Green]] || [[ Filter Set AmR| AmR]] || [[O2k_signal_and_output|Type II]] || 100 - 500 || 1000 || at c(AmR) = 10 µM | | [[Amplex UltraRed|Amplex<sup>TM</sup> UltraRed]]||[[Fluorescence-Sensor Green]] or [[Smart Fluo-Sensor Green]] || [[ Filter Set AmR| AmR]] || [[O2k_signal_and_output|Type II]] || 100 - 500 || 1000 || at c(AmR) = 10 µM | ||
|- | |- | ||
| [[TMRM]]||[[Fluorescence-Sensor Green]]or [[Smart Fluo-Sensor Green]] || [[ Filter Set AmR| AmR]] || [[O2k_signal_and_output|Type III]] || 100 - 500|| 1000 || at c(TMRM) = 2 µM | | [[TMRM]]||[[Fluorescence-Sensor Green]] or [[Smart Fluo-Sensor Green]] || [[ Filter Set AmR| AmR]] || [[O2k_signal_and_output|Type III]] || 100 - 500|| 1000 || at c(TMRM) = 2 µM | ||
|- | |- | ||
| [[Safranin]] ||[[Fluorescence-Sensor Blue]] or [[Smart Fluo-Sensor Blue]]|| [[Filter Set Saf|Saf]] || [[O2k_signal_and_output| Type III]] || 100 - 250 || 1000 || at c(safranin) = 2 µM | | [[Safranin]] ||[[Fluorescence-Sensor Blue]] or [[Smart Fluo-Sensor Blue]]|| [[Filter Set Saf|Saf]] || [[O2k_signal_and_output| Type III]] || 100 - 250 || 1000 || at c(safranin) = 2 µM | ||
Latest revision as of 13:12, 19 November 2021
- 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.
- 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.
| 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 |
