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FRETcalc plugin for calculation of FRET in non-continuous intracellular compartments.

Abstract：

:FRET has emerged as an important tool for studying intracellular processes and interactions between biomolecules. Intracellular donor and acceptor molecules are distributed in individual organelles that usually have complex non-continuous shape. Consequently, background pixels arising from fluorophore-free regions of the cell are proximal to FRET-positive pixels, leading to systemic errors in the estimated FRET values. This study introduces a new FRET(TH) algorithm for FRET estimation by acceptor photobleaching that separates the FRET-positive pixels from the background by applying user-defined thresholds for pixel selection. The FRET(TH) algorithm was validated by analysis of interactions between fluorescently tagged proteins in the endoplasmic reticulum using acquired and simulated images. The novel algorithm showed superior performance to the regular FRET calculation algorithm in acquired images and in most simulations. The developed algorithm was incorporated into the FRETcalc plugin for ImageJ program that enables user-defined choices of thresholds for calculation of FRET by acceptor photobleaching.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Stepensky D

doi

10.1016/j.bbrc.2007.05.180

subject

Has Abstract

pub_date

2007-08-03 00:00:00

pages

752-8

issue

3

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(07)01169-2

journal_volume

359

pub_type

杂志文章

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