FociCounter: a freely available PC programme for quantitative and qualitative analysis of gamma-H2AX foci.
Gamma-H2AX foci are sensitive and specific indicator for the induction of DNA double-strand breaks (DSBs) and an immunocytochemical assay with antibodies recognizing gamma-H2AX has become the gold standard for the detection of this type of DNA lesion. Quantification of gamma-H2AX foci can be achieved by various methods such as Western blotting, flow cytometry, visual analysis and computational analysis with a fluorescence microscope. The best sensitivity is achieved by computer analysis. Since no freeware programme for the analysis of gamma-H2AX foci exists for a PC platform, the aim of our study was to develop a simple and user-friendly public-domain software. The algorithm applied in our programme allows determination of the number of foci in a single cell, a focus intensity per cell, as well as a cell intensity. Its graphical user interface is based on a GTK+ library and the whole application can be run under a variety of operating systems, including MS Windows and Linux. The programme called FociCounter is publicly available at http://focicounter.sourceforge.net. Application of the programme was tested by analysing gamma-H2AX foci in CHO and MO59K cells irradiated in vitro with X-rays and validated by comparing the results obtained with the outcome of automated image analysis and flow cytometry.
Jucha
Anna
A
Institute of Theoretical Physics, University of Wroclaw, Poland.
Wegierek-Ciuk
Aneta
A
Koza
Zbigniew
Z
Lisowska
Halina
H
Wojcik
Andrzej
A
Wojewodzka
Maria
M
Lankoff
Anna
A
D000465:Algorithms; D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D003201:Computers; D006224:Cricetinae; D003412:Cricetulus; D053903:DNA Breaks, Double-Stranded; D006657:Histones / Q000235:genetics*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D011829:Radiation Dosage; D011832:Radiation Injuries; D012984:Software; D014965:X-Rays
C468783:H2AX protein, human; D006657:Histones