Monitoring dynamic binding of chromatin proteins in vivo by single-molecule tracking

Davide Mazza, Sourav Ganguly, James G. McNally

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Single-molecule fluorescence microscopy has been used for decades to quantify macromolecular dynamics occurring in specimens that are in direct contact with a coverslip. This has permitted in vitro analysis of single-molecule motion in various biochemically reconstituted systems as well as in vivo studies of single-molecule motion on cell membranes. More recently, thanks to improvements in fluorescent tags and microscopes, it has been possible to follow individual molecules inside thicker specimens such as the nucleus of living cells. This has enabled a detailed description of the live-cell binding of nuclear proteins to DNA. In this protocol we describe a method to quantify intranuclear binding using single-molecule tracking (SMT).

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages117-137
Number of pages21
Volume1042
ISBN (Print)9781627035255
DOIs
Publication statusPublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1042
ISSN (Print)10643745

Fingerprint

Chromatin
Carrier Proteins
Nuclear Proteins
Cell Nucleus
Fluorescence Microscopy
Cell Membrane
DNA
Single Molecule Imaging
In Vitro Techniques

Keywords

  • DNA binding
  • Microscopy
  • Single-molecule tracking
  • Transcription factor

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Mazza, D., Ganguly, S., & McNally, J. G. (2013). Monitoring dynamic binding of chromatin proteins in vivo by single-molecule tracking. In Methods in Molecular Biology (Vol. 1042, pp. 117-137). (Methods in Molecular Biology; Vol. 1042). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-526-2_9

Monitoring dynamic binding of chromatin proteins in vivo by single-molecule tracking. / Mazza, Davide; Ganguly, Sourav; McNally, James G.

Methods in Molecular Biology. Vol. 1042 Humana Press Inc., 2013. p. 117-137 (Methods in Molecular Biology; Vol. 1042).

Research output: Chapter in Book/Report/Conference proceedingChapter

Mazza, D, Ganguly, S & McNally, JG 2013, Monitoring dynamic binding of chromatin proteins in vivo by single-molecule tracking. in Methods in Molecular Biology. vol. 1042, Methods in Molecular Biology, vol. 1042, Humana Press Inc., pp. 117-137. https://doi.org/10.1007/978-1-62703-526-2_9
Mazza D, Ganguly S, McNally JG. Monitoring dynamic binding of chromatin proteins in vivo by single-molecule tracking. In Methods in Molecular Biology. Vol. 1042. Humana Press Inc. 2013. p. 117-137. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-526-2_9
Mazza, Davide ; Ganguly, Sourav ; McNally, James G. / Monitoring dynamic binding of chromatin proteins in vivo by single-molecule tracking. Methods in Molecular Biology. Vol. 1042 Humana Press Inc., 2013. pp. 117-137 (Methods in Molecular Biology).
@inbook{77eb83f8390a4d9d9b5d5946f58ba319,
title = "Monitoring dynamic binding of chromatin proteins in vivo by single-molecule tracking",
abstract = "Single-molecule fluorescence microscopy has been used for decades to quantify macromolecular dynamics occurring in specimens that are in direct contact with a coverslip. This has permitted in vitro analysis of single-molecule motion in various biochemically reconstituted systems as well as in vivo studies of single-molecule motion on cell membranes. More recently, thanks to improvements in fluorescent tags and microscopes, it has been possible to follow individual molecules inside thicker specimens such as the nucleus of living cells. This has enabled a detailed description of the live-cell binding of nuclear proteins to DNA. In this protocol we describe a method to quantify intranuclear binding using single-molecule tracking (SMT).",
keywords = "DNA binding, Microscopy, Single-molecule tracking, Transcription factor",
author = "Davide Mazza and Sourav Ganguly and McNally, {James G.}",
year = "2013",
doi = "10.1007/978-1-62703-526-2_9",
language = "English",
isbn = "9781627035255",
volume = "1042",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "117--137",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Monitoring dynamic binding of chromatin proteins in vivo by single-molecule tracking

AU - Mazza, Davide

AU - Ganguly, Sourav

AU - McNally, James G.

PY - 2013

Y1 - 2013

N2 - Single-molecule fluorescence microscopy has been used for decades to quantify macromolecular dynamics occurring in specimens that are in direct contact with a coverslip. This has permitted in vitro analysis of single-molecule motion in various biochemically reconstituted systems as well as in vivo studies of single-molecule motion on cell membranes. More recently, thanks to improvements in fluorescent tags and microscopes, it has been possible to follow individual molecules inside thicker specimens such as the nucleus of living cells. This has enabled a detailed description of the live-cell binding of nuclear proteins to DNA. In this protocol we describe a method to quantify intranuclear binding using single-molecule tracking (SMT).

AB - Single-molecule fluorescence microscopy has been used for decades to quantify macromolecular dynamics occurring in specimens that are in direct contact with a coverslip. This has permitted in vitro analysis of single-molecule motion in various biochemically reconstituted systems as well as in vivo studies of single-molecule motion on cell membranes. More recently, thanks to improvements in fluorescent tags and microscopes, it has been possible to follow individual molecules inside thicker specimens such as the nucleus of living cells. This has enabled a detailed description of the live-cell binding of nuclear proteins to DNA. In this protocol we describe a method to quantify intranuclear binding using single-molecule tracking (SMT).

KW - DNA binding

KW - Microscopy

KW - Single-molecule tracking

KW - Transcription factor

UR - http://www.scopus.com/inward/record.url?scp=84934434809&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84934434809&partnerID=8YFLogxK

U2 - 10.1007/978-1-62703-526-2_9

DO - 10.1007/978-1-62703-526-2_9

M3 - Chapter

C2 - 23980004

AN - SCOPUS:84934434809

SN - 9781627035255

VL - 1042

T3 - Methods in Molecular Biology

SP - 117

EP - 137

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -