Differences

This shows you the differences between two versions of the page.

--- howto:calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script [2014/06/17 15:36] – admin
+++ howto:calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script [2014/06/18 16:06] – admin
@@ Line 1: / Line 1: @@
 {{tag> tutorial howto FRET PIE analysis time-trace SymPhoTime}}
-~~TOC~~
+~~NOTOC~~
-====== Calculate Ratiometric Single Pair FRET Distributions Using the PIE-FRET Analysis ======
+====== Calculate Ratiometric Single Pair FRET Distributions Using PIE-FRET ======
@@ Line 10: / Line 10: @@
 This tutorial shows step-by-step, how to calculate a FRET histogram from single [[glossary:fret|FRET]]pairs detected under single molecule conditions using pulsed interleaved excitation (PIE). The script requires a spectrally resolved fluorescence time trace, in which donor and acceptor have been excited alternately. As a result, the distribution of FRET efficiencies of different molecules is shown which can be used to detect subpopulations.
+{{ youtube>cSAbgJpZhrY?large }}
 **Note:** The script requires a time trace containing the fluorescence of two spectrally separated channels, one channel mainly detecting the fluorescence of the donor dye and the other channel detecting mainly the fluorescence of the acceptor dye.\\
@@ Line 162: / Line 164: @@
 **Note:** Further information about the sample used in this measurement can be found under: J.L. Fiore et. al., Enthalpy-driven RNA-folding: single-molecule thermodynamics of tetraloop-receptor tertiary interaction,  Biochemistry (2009), 48(11), 2550-8. ([[http://pubs.acs.org/doi/abs/10.1021/bi8019788]]).