Authors
- János Roszik - janosr@med.unideb.hu,
- János Szöllõsi - szollo@med.unideb.hu and
- György Vereb - vereb@med.unideb.hu, Department of Biophysics and Cell Biology, Research Center for Molecular Medicine,University of Debrecen, Debrecen, Hungary
Please send questions, comments and requests about the program to János Roszik.
Description
AccPbFRET is an ImageJ plugin which calculates FRET (fluorescence resonance energy transfer) efficiencies pixel-by-pixel based on images taken by this microscopy technique. It allows the user to correct for the donor photobleaching, acceptor cross-talk, acceptor photoproduct and partial acceptor photobleaching effects. Furthermore, automatic registration and semi-automatic analysis of large image sets is also provided in this software tool.
If you are using the plugin, please cite the following paper, in which the program and the analysis are described:
Roszik J, Szollosi J, Vereb G: AccPbFRET: an ImageJ plugin for semi-automatic, fully corrected analysis of acceptor photobleaching FRET images BMC Bioinformatics 2008, 9:346
(It can be downloaded from the homepage of BMC Bioinformatics at BioMed Central.)
Prospective users should read this paper before running the program.
Terms of Use
- You can download and use the program for your data analysis freely.
- You may modify the program for your own personal usage.
Download (version 3.15, 26 Mar 2009)
- AccPbFRET_Plugin.java (Java source code)
Requirements
- a computer that runs ImageJ 1.42k (with Java 1.6.0_05) /other versions of 1.42 or higher should also work, but have not been tested with this version/
(Version 1.42k ij.jar can be downloaded from here. ImageJ can be easily updated with: Plugins->Utilities->Update ImageJ...) - screen resolution 1280 x 900 or higher
(Java window elements can have different size on different platforms/computers. If you cannot see the buttons correctly, please try to increase the window width/height.)
Installation
- Download AccPbFRET_Plugin.java to IMAGEJ_HOME_DIRECTORY \ plugins
- select the menu "Plugins" in ImageJ and choose "Compile and Run..."
- select AccPbFRET_Plugin.java and click "Open"
Download example images: (cell line: SK-BR-3, donor: Alexa-555-Herceptin, acceptor: Cy5-2C4)
The program works with single tiff images, and can handle lsm image stacks as well.
- sample with donor and acceptor: accpbfret-example-images-donor-acceptor.lsm
- sample with donor only: accpbfret-example-images-donor-only.lsm
- sample with acceptor only: accpbfret-example-images-acceptor-only.lsm
- some example lsm files for semi-automatic processing: accpbfret-semiautomatic-example.zip
For live cell imaging, we recommend our other FRET plugin RiFRET.