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Related Experiment Video

Updated: Apr 8, 2026

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins

Published on: September 28, 2012

Characterizing the DNA Damage Response by Cell Tracking Algorithms and Cell Features Classification Using

Walter Georgescu¹, Alma Osseiran¹, Maria Rojec¹

¹Lawrence Berkeley National Laboratory, Life Sciences Division, Berkeley, CA, United States of America.

|June 25, 2015

Summary

Live cell imaging reveals DNA repair dynamics, showing radiation-induced foci merge into repair domains. This process, especially at high radiation doses, may explain increased DNA damage and cell death.

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Last Updated: Apr 8, 2026

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins

Published on: September 28, 2012

Laser Microirradiation to Study In Vivo Cellular Responses to Simple and Complex DNA Damage

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Application of Laser Micro-irradiation for Examination of Single and Double Strand Break Repair in Mammalian Cells

Application of Laser Micro-irradiation for Examination of Single and Double Strand Break Repair in Mammalian Cells

Published on: September 5, 2017

Area of Science:

Cell Biology
Molecular Biology
Biophysics

Background:

Traditional DNA repair kinetics studies use fixed cells, limiting dynamic analysis.
Understanding DNA damage response (DDR) dynamics across cell generations is crucial.

Purpose of the Study:

To characterize the dynamics of 53BP1 radiation-induced foci (RIF) across multiple cell generations using live cell imaging.
To quantify the dose-dependent behavior of RIFs after ionizing radiation exposure.

Main Methods:

Live cell imaging of MCF10A cells expressing histone H2B-GFP and 53BP1-mCherry.
Automatic extraction of RIF imaging features and linear programming for kinetic analysis.
Elastic registration for precise RIF motion tracking within the nucleus.

Main Results:

RIFs exhibit random motion and diffusion within repair domains, with merging being the primary mode of large foci formation.
Repair domains were estimated at 7.5–11 µm², with ~15 per cell.
High radiation doses (>1 Gy) induced rapid 53BP1 increase and faster foci diffusion, suggesting a stressed repair process.

Conclusions:

RIF merging at high doses may increase double-strand break (DSB) proximity, leading to misrepair.
This mechanism could explain the supralinear dose dependence of chromosomal rearrangements and cell death after ionizing radiation.