Computer program for determining fluorescence resonance energy transfer efficiency from flow cytometric data on a cell-by-cell basis

Gergely Szentesi, Gábor Horváth, Imre Bori, G. Vámosi, J. Szöllősi, R. Gáspár, S. Damjanovich, A. Jenei, L. Mátyus

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

The determination of fluorescence resonance energy transfer (FRET) with flow cytometry (FCET) is one of the most efficient tools to study the proximity relationships of cell membrane components in cell populations on a cell-by-cell basis. Because of the high amount of data and the relatively tedious calculations, this procedure should be assisted by powerful data processing software. The currently available programs are not able to fulfill this requirement. We developed a Windows-based program to calculate fluorescence resonance energy transfer efficiency values from list mode flow cytometry standard (FCS) files. This program displays the measured data in standard plots by generating one- and two-parameter histograms on linear or logarithmic scales. A graphical gating tool allows the user to select the desired cell population according to any combination of the parameter values. The program performs several statistical calculations, including mean, S.D., percent of the gated data. We have implemented two types of data sheet for FRET calculations to aid and guide the user during the analysis: one with population-mean-based autofluorescence correction and the other with spectrum-based cell-by-cell autofluorescence correction. In this paper, we describe the gating algorithms, the file opening procedure and the rules of gating. The structure of the program and a short description of the graphical user-interface (GUI) are also presented in this article.

Original languageEnglish
Pages (from-to)201-211
Number of pages11
JournalComputer Methods and Programs in Biomedicine
Volume75
Issue number3
DOIs
Publication statusPublished - Sep 2004

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Fluorescence Resonance Energy Transfer
Computer program listings
Software
Flow cytometry
Efficiency
Cells
Cell membranes
Graphical user interfaces
Flow Cytometry
Population
Data Display
Cellular Structures
Cell Membrane

Keywords

  • Computer program
  • Flow cytometry
  • Fluorescence resonance energy transfer
  • Gating algorithm

ASJC Scopus subject areas

  • Software

Cite this

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abstract = "The determination of fluorescence resonance energy transfer (FRET) with flow cytometry (FCET) is one of the most efficient tools to study the proximity relationships of cell membrane components in cell populations on a cell-by-cell basis. Because of the high amount of data and the relatively tedious calculations, this procedure should be assisted by powerful data processing software. The currently available programs are not able to fulfill this requirement. We developed a Windows-based program to calculate fluorescence resonance energy transfer efficiency values from list mode flow cytometry standard (FCS) files. This program displays the measured data in standard plots by generating one- and two-parameter histograms on linear or logarithmic scales. A graphical gating tool allows the user to select the desired cell population according to any combination of the parameter values. The program performs several statistical calculations, including mean, S.D., percent of the gated data. We have implemented two types of data sheet for FRET calculations to aid and guide the user during the analysis: one with population-mean-based autofluorescence correction and the other with spectrum-based cell-by-cell autofluorescence correction. In this paper, we describe the gating algorithms, the file opening procedure and the rules of gating. The structure of the program and a short description of the graphical user-interface (GUI) are also presented in this article.",
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AU - Horváth, Gábor

AU - Bori, Imre

AU - Vámosi, G.

AU - Szöllősi, J.

AU - Gáspár, R.

AU - Damjanovich, S.

AU - Jenei, A.

AU - Mátyus, L.

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