Labelling by immunofluorescence of gamma-H2AX acquired with confocal microscopy.

<p>This allows the study of the distribution of foci in the whole volume of irradiated endothelial cell nuclei, cultured as monolayers on glass in Lab-Tek^® chambers. (A) Gamma-H2AX foci viewed at the focal plane, as for conventional microscopy. (B) Top view after 3D reconstruction, with confocal microscopy acquisition of 15 layers across the nucleus. The number of foci is identical to that observed at the focal plane. (C) Side view of the 3D reconstruction. In view of the size of the foci, the thickness of the endothelial nucleus and the depth of field of the x63 objective used, it appears that all foci that can be detected with confocal microscopy can also be detected at focal plane with conventional microscopy. The picture is representative of all the nuclei examined by confocal microscopy.