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.
Protocol for induction and study of DNA double-strand breaks in mammalian cells using PALM microdissection and expansion microscopy.
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作者：Luna-Maldonado Fernando, Oliva-Rico Diego, López-Saavedra Alejandro, Arenas-Hernández Aquetzalli, Castro-Hernández Clementina, Rodriguez Gatica Juan Eduardo, Pérez-Yepez Eloy-Andres, DÃaz-Chávez José, Herrera Luis A
| 期刊： | STAR Protocols | 影响因子： | 1.300 |
| 时间： | 2025 | 起止号： | 2025 Jul 9; 6(3):103938 |
| doi： | 10.1016/j.xpro.2025.103938 | ||
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