High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation

Scientific Reports
Sebastian OeckPeter M Glazer

Abstract

DNA- and histone-related research frequently comprises the quantitative analysis of protein modifications, such as histone phosphorylation. Analysis of accumulation and disappearance of protein foci are used to monitor DNA damage and repair kinetics. If the protein of interest doesn't accumulate in foci, laser micro-irradiation of single nuclei provides an alternative method to monitor DNA repair proteins and histone dynamics at the DNA damage site. We have developed an automated evaluation tool for standardized, high-throughput analysis of micro-irradiated cells featuring single cell background subtraction and detection across multiple fluorescence channels, allowing for robust statistics.

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Citations

Oct 7, 2020·International Journal of Molecular Sciences·Sebastian OeckAlexander Schramm
Oct 28, 2019·Journal of Controlled Release : Official Journal of the Controlled Release Society·Hanna K MandlW Mark Saltzman

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Methods Mentioned

BETA
fluorescence microscopy
acetylation
irradiate
Fluorescence

Software Mentioned

ImageJ Bio - Formats
Windows
Watershed
Excel
Plot Profile
ImageJ Plot
Focinator
NIS Elements
Prism8
ImageJ

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