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Updated: Jul 14, 2026
Analysis of DNA Double-strand Break DSB Repair in Mammalian Cells
Published on: September 8, 2010
Protocol for induction and study of DNA double-strand breaks in mammalian cells using PALM microdissection and
Fernando Luna-Maldonado1, Diego Oliva-Rico2, Alejandro López-Saavedra3
1Unidad de Investigación Biomédica en Cáncer, Instituto Nacional de Cancerología (INCan)- Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México (UNAM), Mexico City 14080, Mexico; Instituto Nacional de Cancerología, Advanced Microscopy Applications Unit (ADMiRA), Av. San Fernando 22, Col. Sección XVI, Tlalpan 14080, CDMX, México.
Abstract:
We present a protocol to induce localized DNA double-strand breaks (DSBs) in mammalian monolayer cells, using the PALM MicroBeam microdissector, followed by high-resolution visualization through expansion microscopy (ExM). We describe steps for laser microirradiation, immunostaining, and gel embedding. The protocol enables 4.5-fold physical expansion and confocal imaging of DNA damage sites. We quantify nuclear and damage site expansion using Fiji by measuring nuclear area and cross-sectional fluorescence profiles.
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