howto:flim_fret_calculation_for_multi_exponential_donors
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howto:flim_fret_calculation_for_multi_exponential_donors [2014/05/23 07:43] – admin | howto:flim_fret_calculation_for_multi_exponential_donors [2020/05/12 12:37] – [Step-by-Step Tutorial] ruckelshausen | ||
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====== FLIM FRET Calculation for Multi Exponential Donors ====== | ====== FLIM FRET Calculation for Multi Exponential Donors ====== | ||
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This tutorial shows step-by-step, | This tutorial shows step-by-step, | ||
- | **Note:**\\ | + | {{ youtube> |
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+ | ===== Background information ===== | ||
FLIM-FRET is a robust method to determine the FRET efficiency of a suited donor acceptor pair. | FLIM-FRET is a robust method to determine the FRET efficiency of a suited donor acceptor pair. | ||
If the FRET-donor molecule has a more exponential decay kinetic, the amplitude weighted average lifetime has to be used to calculate the average FRET efficiency. | If the FRET-donor molecule has a more exponential decay kinetic, the amplitude weighted average lifetime has to be used to calculate the average FRET efficiency. | ||
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===== Step-by-Step Tutorial ===== | ===== Step-by-Step Tutorial ===== | ||
+ | **Note:** This Step-by-Step tutorial was written for SymPhoTime version 2.5. There might be slight differences compared to other software versions. For example the way of setting an intensity threshold has changed. In the video the former version is shown. | ||
==== Determine the donor only lifetime using the FLIM script ==== | ==== Determine the donor only lifetime using the FLIM script ==== | ||
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* To analyze many images with the same FRET pair, it is possible to save the starting values for the fits by clicking on "Save Defaults" | * To analyze many images with the same FRET pair, it is possible to save the starting values for the fits by clicking on "Save Defaults" | ||
* If you don't want to do anything more, just close the result window. | * If you don't want to do anything more, just close the result window. | ||
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howto/flim_fret_calculation_for_multi_exponential_donors.txt · Last modified: 2023/11/21 10:33 by lan