Immunofluorescence-based imaging can offer high signal-to-noise and overall superior image quality of fixed cells over that of live-cell imaging. This is especially true of tracking and quantifying DNA damage sites as there is a dearth of reliable or easy-to-use methods in living cells. Here, I describe a useful workflow for DNA damage induction in cell culture, fixation, staining, and method for analyzing DNA double-strand breaks (DSBs). Further, I provide troubleshooting tips for common issues while performing image segmentation and analysis.

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Imaging Approach to DNA Damage Induction and Quantification

  • Mikael V. Garabedian

摘要

Immunofluorescence-based imaging can offer high signal-to-noise and overall superior image quality of fixed cells over that of live-cell imaging. This is especially true of tracking and quantifying DNA damage sites as there is a dearth of reliable or easy-to-use methods in living cells. Here, I describe a useful workflow for DNA damage induction in cell culture, fixation, staining, and method for analyzing DNA double-strand breaks (DSBs). Further, I provide troubleshooting tips for common issues while performing image segmentation and analysis.