Monitoring dynamic binding of chromatin proteins in vivo by fluorescence correlation spectroscopy and temporal image correlation spectroscopy

Davide Mazza, Timothy J. Stasevich, Tatiana S. Karpova, James G. McNally

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Live-cell microscopy has demonstrated that many nuclear proteins bind transiently to target sites in chromatin. These binding interactions can be detected and quantified by two related live-cell imaging techniques, Fluorescence Correlation Spectroscopy (FCS) and Temporal Image Correlation Spectroscopy (TICS). With proper quantitative modeling, it is possible to obtain estimates from FCS and TICS data of the association and dissociation rates of nuclear protein binding to chromatin. These binding rates permit calculating the fractions of free and bound protein in the nucleus, plus the time required to diffuse from one binding site to the next and the dwell time on a chromatin target. In this protocol, we summarize the underlying principles of FCS and TICS, and then describe how these data should be collected and analyzed to extract estimates of in vivo binding.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Pages177-200
Number of pages24
Volume833
DOIs
Publication statusPublished - 2012

Publication series

NameMethods in Molecular Biology
Volume833
ISSN (Print)10643745

Keywords

  • Binding
  • Fluorescence correlation spectroscopy
  • In vivo
  • Nuclear protein
  • Temporal image correlation spectroscopy

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Monitoring dynamic binding of chromatin proteins in vivo by fluorescence correlation spectroscopy and temporal image correlation spectroscopy'. Together they form a unique fingerprint.

  • Cite this

    Mazza, D., Stasevich, T. J., Karpova, T. S., & McNally, J. G. (2012). Monitoring dynamic binding of chromatin proteins in vivo by fluorescence correlation spectroscopy and temporal image correlation spectroscopy. In Methods in Molecular Biology (Vol. 833, pp. 177-200). (Methods in Molecular Biology; Vol. 833). https://doi.org/10.1007/978-1-61779-477-3_12