Immunofluorescence microscopy-based assessment of cytosolic DNA accumulation in mammalian cells

Ai Sato, Aitziber Buque, Takahiro Yamazaki, Norma Bloy, Giulia Petroni, Lorenzo Galluzzi

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Here, we describe an immunofluorescence (IF) microscopy-based approach to quantify cytosolic double-stranded DNA molecules in cultured eukaryotic cells upon the selective and specific permeabilization of plasma membranes. This technique is compatible with widefield microscopy coupled with automated image analysis for mid- to high-throughput applications and high-resolution confocal microscopy for subcellular assessments and co-localization studies. In addition to enabling single-cell and subcellular resolution, this approach circumvents most constraints associated with alternative approaches based on subcellular fractionation. For complete use and execution of this protocol, please refer to Yamazaki et al. (2020).

Original languageEnglish
Article number100488
JournalStar Protocols
Volume2
Issue number2
DOIs
StatePublished - Jun 18 2021
Externally publishedYes

Keywords

  • Antibody
  • Cancer
  • Cell culture
  • Cell-based Assays
  • Immunology
  • Microscopy

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