title: In-solution staining and arraying method for the immunofluorescence detection of γH2AX foci optimized for clinical applications.
abstract: Immunofluorescence quantification of γH2AX foci is a powerful approach to quantify DNA double-strand breaks induced by cancer therapy or accidental exposure to ionizing radiation. Here we report a modification to the γH2AX immunofluorescence labeling method, whereby cells are stained in-solution before being spotted and fixed onto microscope slides. Our modified method allows arraying of 16 patient samples/slide ready for foci counting in 2 h and demonstrated reliably detection of γH2AX foci in mononuclear cells prepared from patients who had undergone radiation therapy.
authors: Department of Clinical Chemistry and Transfusion Medicine, The Institute of Biomedicine, The Sahlgrenska Academy at University of Gothenburg, Sweden.
authors: Pegah
authors: P
authors: Johansson
authors: Aida
authors: A
authors: Muslimovic
authors: Ragnar
authors: R
authors: Hultborn
authors: Erik
authors: E
authors: Fernström
authors: Ola
authors: O
authors: Hammarsten
mesh_terms: D000617:Aminoglycosides / Q000494:pharmacology; D053903:DNA Breaks, Double-Stranded; D053281:Enediynes / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006657:Histones / Q000032:analysis*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry* / Q000187:drug effects / Q000528:radiation effects; D009369:Neoplasms / Q000532:radiotherapy*; D013194:Staining and Labeling / Q000379:methods; D046888:Tissue Array Analysis / Q000379:methods*
chemical_list: D000617:Aminoglycosides; D053281:Enediynes; C468783:H2AX protein, human; D006657:Histones; C074528:calicheamicin T