AccPbFRET: An ImageJ plugin for semi-automatic, fully corrected analysis of acceptor photobleaching FRET imagesReport as inadecuate




AccPbFRET: An ImageJ plugin for semi-automatic, fully corrected analysis of acceptor photobleaching FRET images - Download this document for free, or read online. Document in PDF available to download.

BMC Bioinformatics

, 9:346

First Online: 19 August 2008Received: 23 April 2008Accepted: 19 August 2008

Abstract

BackgroundThe acceptor photobleaching fluorescence resonance energy transfer FRET method is widely used for monitoring molecular interactions in cells. This method of FRET, while among those with the simplest mathematics, is robust, self-controlled and independent of fluorophore amounts and ratios.

ResultsAccPbFRET is a user-friendly, efficient ImageJ plugin which allows fully corrected, pixel-wise calculation and detailed, ROI region of interest-based analysis of FRET efficiencies in microscopic images. Furthermore, automatic registration and semi-automatic analysis of large image sets is provided, which are not available in any existing FRET evaluation software.

ConclusionDespite of the widespread applicability of the acceptor photobleaching FRET technique, this is the first paper where all possible sources of major errors of the measurement and analysis are considered, and AccPbFRET is the only program which provides the complete suite of corrections – for registering image pairs, for unwanted photobleaching of the donor, for cross-talk of the acceptor and-or its photoproduct to the donor channel and for partial photobleaching of the acceptor. The program efficiently speeds up the analysis of large image sets even for novice users and is freely available.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2105-9-346 contains supplementary material, which is available to authorized users.

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Author: János Roszik - János Szöllősi - György Vereb

Source: https://link.springer.com/







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