Protocol for induction and study of DNA double-strand breaks in mammalian cells using PALM microdissection and expansion microscopy

Protocol for induction and study of DNA double-strand breaks in mammalian cells using PALM microdissection and expansion microscopy

Summary: 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 ge...

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Bibliographic Details
Main Authors: Fernando Luna-Maldonado, Diego Oliva-Rico, Alejandro López-Saavedra, Aquetzalli Arenas-Hernández, Clementina Castro-Hernández, Juan Eduardo Rodriguez Gatica, Eloy-Andres Pérez-Yepez, José Díaz-Chávez, Luis A. Herrera
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:STAR Protocols
Subjects:
Cell Biology
Cancer
Microscopy
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725003442
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Summary:Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667

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