The quantitative detection of radiation caused DNA double-strand breaks (DSB) by immunostained \u03b3-H2AX foci using direct stochastic optical reconstruction microscopy (dSTORM) provides a deeper insight into the DNA repair process at nanoscale in a time-dependent manner. <\/p>\n\n\n\n\n\n\n\n
Glioblastoma (U251) cells were irradiated with 250 keV X-ray at 0, 2, 5, 8 Gy dose levels. Cell cycle phase distribution and apoptosis of U251 cells upon irradiation was assayed by flow cytometry. We studied the density, topology and volume of the \u03b3-H2AX foci with 3D confocal microscopy and the dSTORM superresolution method. A pronounced increase in \u03b3-H2AX foci and cluster density was detected by 3D confocal microscopy after 2 Gy, at 30 min postirradiation, but both returned to the control level at 24 h. Meanwhile, at 24 h a considerable amount of residual foci could be measured from 5 Gy, which returned to the normal level 48 h later. The dSTORM based \u03b3-H2AX analysis revealed that the micron-sized \u03b3-H2AX foci are composed of distinct smaller units with a few tens of nanometers. The density of these clusters, the epitope number and the dynamics of \u03b3-H2AX foci loss could be analyzed. Our findings suggest a discrete level of repair enzyme capacity and the restart of the repair process for the residual DSBs, even beyond 24 h. The dSTORM superresolution technique provides a higher precision over 3D confocal microscopy to study radiation induced \u03b3-H2AX foci and molecular rearrangements during the repair process, opening a novel perspective for radiation research. https:\/\/doi.org\/10.3389\/pore.2021.1609971<\/a><\/p>\n","protected":false},"excerpt":{"rendered":" The quantitative detection of radiation caused DNA double-strand breaks (DSB) by immunostained \u03b3-H2AX foci using direct stochastic optical reconstruction microscopy (dSTORM) provides a deeper insight into the DNA repair process<\/p>\n","protected":false},"author":3,"featured_media":1764,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_publicize_message":"","jetpack_publicize_feature_enabled":true,"jetpack_social_post_already_shared":true,"jetpack_social_options":{"image_generator_settings":{"template":"highway","default_image_id":0,"font":"","enabled":false},"version":2},"jetpack_post_was_ever_published":false},"categories":[10],"tags":[17,23],"class_list":["post-1763","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-publication","tag-press","tag-publication"],"jetpack_publicize_connections":[],"jetpack_featured_media_url":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/wp-content\/uploads\/2021\/11\/por_marketting_log.jpg","jetpack_sharing_enabled":true,"_links":{"self":[{"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=\/wp\/v2\/posts\/1763","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=1763"}],"version-history":[{"count":2,"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=\/wp\/v2\/posts\/1763\/revisions"}],"predecessor-version":[{"id":1766,"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=\/wp\/v2\/posts\/1763\/revisions\/1766"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=\/wp\/v2\/media\/1764"}],"wp:attachment":[{"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1763"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1763"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/titan.physx.u-szeged.hu\/~adoptim\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1763"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}