Three-color dSTORM Imaging and Analysis of Recombination Foci in Mouse Spread Meiotic Nuclei.

ABSTRACT: During the first meiotic prophase in mouse, repair of SPO11-induced DNA double-strand breaks (DSBs), facilitating homologous chromosome synapsis, is essential to successfully complete the first meiotic cell division. Recombinases RAD51 and DMC1 play an important role in homology search, but their mechanistic contribution to this process is not fully understood. Super-resolution, single-molecule imaging of RAD51 and DMC1 provides detailed information on recombinase accumulation on DSBs during meiotic prophase. Here, we present a detailed protocol of recombination foci analysis of three-color direct stochastic optical reconstruction microscopy (dSTORM) imaging of SYCP3, RAD51, and DMC1, fluorescently labeled by antibody staining in mouse spermatocytes. This protocol consists of sample prepar

SUBMITTER: Koornneef L 

PROVIDER: S-EPMC10367009 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature