4.3 Article

In-solution staining and arraying method for the immunofluorescence detection of γH2AX foci optimized for clinical applications

Journal

BIOTECHNIQUES
Volume 51, Issue 3, Pages 185-189

Publisher

BIOTECHNIQUES OFFICE
DOI: 10.2144/000113738

Keywords

H2AX; gamma H2AX; H2AX-foci; cancer; irradiation; immunofluorescence staining; spotting/arraying

Funding

  1. Swedish Cancer Society
  2. Swedish Research Council
  3. Sahlgrenska University Hospital
  4. Swedish Pain Foundation (SSF)
  5. King Gustav V Jubilee Clinic Cancer Research Foundation
  6. Assar Gabrielsson Cancer Research Foundation

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Immunofluorescence quantification of gamma H2AX 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 gamma H2AX 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 gamma H2AX foci in mononuclear cells prepared from patients who had undergone radiation therapy.

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