![]() Several publications showed that manual counting of foci is time consuming, frequently inaccurate and subjected to investigator-related bias. However, the programs currently available for counting and analysis of nuclei are often based on manual analysis. To evaluate formation and processing of these DNA damage foci, a reliable and accurate image analysis is required.ĭue to the wide use of methods retrieving images of foci and cells, multiple evaluation procedures have been developed. This method takes advantage of the fact that many repair proteins and repair associated proteins, such as γ-H2.AX, 53BP1 and RAD51, accumulate and co-localize at the site of DNA damage. Another possibility is to fuse the proteins of interest with fluorescent proteins, such as green fluorescence protein GFP. phospho-histone 2.AX (γ-H2.AX)) antibodies directly linked to a fluorophore or detected by using a secondary fluorophore-labeled antibody. p53 binding protein 1 (53BP1)) or phospho-protein-specific (e.g. For this purpose the proteins of interest or their specific phosphorylated isoform are visualized by immunofluorescence using protein-specific (e.g. To evaluate the efficacy of IR alone or in combination with chemotherapy or drugs inducing DNA damage and targeting DNA repair, radiation biologists usually count fluorescence-labeled protein-foci in the nucleus using fluorescence microscopy. Radiotherapy (RT) is a mainstay in modern cancer treatment. ![]()
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