Full Content is available to subscribers

Subscribe/Learn More  >
Proceedings Article

Quantification of DNA repair protein kinetics after γ-irradiation using number and brightness analysis

[+] Author Affiliations
Salim Abdisalaam, George Alexandrakis

The Univ. of Texas at Arlington (USA)

Milan Poudel, David J. Chen

The Univ. of Texas Southwestern Medical Ctr. at Dallas (USA)

Proc. SPIE 7905, Single Molecule Spectroscopy and Imaging IV, 790517 (February 22, 2011); doi:10.1117/12.873853
Text Size: A A A
From Conference Volume 7905

  • Single Molecule Spectroscopy and Imaging IV
  • Jörg Enderlein; Zygmunt K. Gryczynski; Rainer Erdmann
  • San Francisco, California, USA | January 22, 2011

abstract

The kinetics of most proteins involved in DNA damage sensing, signaling and repair following ionizing radiation exposure cannot be quantified by current live cell fluorescence microscopy methods. This is because most of these proteins, with only few notable exceptions, do not attach in large numbers at DNA damage sites to form easily detectable foci in microscopy images. As a result a high fluorescence background from freely moving and immobile fluorescent proteins in the nucleus masks the aggregation of proteins at sparse DNA damage sites. Currently, the kinetics of these repair proteins are studied by laser-induced damage and Fluorescence Recovery After Photobleaching that rely on the detectability of high fluorescence intensity spots of clustered DNA damage. We report on the use of Number and Brightness (N&B) analysis methods as a means to monitor kinetics of DNA repair proteins during sparse DNA damage created by γ-irradiation, which is more relevant to cancer treatment than laser-induced clustered damage. We use two key double strand break repair proteins, namely Ku 70/80 and the DNA-dependent protein kinase catalytic subunit (DNA-PKCS), as specific examples to showcase the feasibility of the proposed methods to quantify dose-dependent kinetics for DNA repair proteins after exposure to γ-rays.

© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Salim Abdisalaam ; Milan Poudel ; David J. Chen and George Alexandrakis
"Quantification of DNA repair protein kinetics after γ-irradiation using number and brightness analysis", Proc. SPIE 7905, Single Molecule Spectroscopy and Imaging IV, 790517 (February 22, 2011); doi:10.1117/12.873853; http://dx.doi.org/10.1117/12.873853


Access This Proceeding
Sign in or Create a personal account to Buy this proceeding ($15 for members, $18 for non-members).

Figures

Tables

NOTE:
Citing articles are presented as examples only. In non-demo SCM6 implementation, integration with CrossRef’s "Cited By" API will populate this tab (http://www.crossref.org/citedby.html).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Advertisement
  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Proceeding
Sign in or Create a personal account to Buy this proceeding ($15 for members, $18 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.