Time-integrated fluorescence cumulant analysis and its application in living cells

Bin Wu, Robert H. Singer, Joachim D. Mueller

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Citations (Scopus)

Abstract

Time-integrated fluorescence cumulant analysis (TIFCA) is a data analysis technique for fluorescence fluctuation spectroscopy (FFS) that extracts information from the cumulants of the integrated fluorescence intensity. It is the first exact theory that describes the effect of sampling time on FFS experiment. Rebinning of data to longer sampling times helps to increase the signal/noise ratio of the experimental cumulants of the photon counts. The sampling time dependence of the cumulants encodes both brightness and diffusion information of the sample. TIFCA analysis extracts this information by fitting the cumulants to model functions. Generalization of TIFCA to multicolor FFS experiment is straightforward. Here, we present an overview of the theory, its implementation, as well as the benefits and requirements of TIFCA. The questions of why, when, and how to use TIFCA will be discussed. We give several examples of practical applications of TIFCA, particularly focused on measuring molecular interaction in living cells.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
Pages99-119
Number of pages21
Volume518
DOIs
StatePublished - 2013

Publication series

NameMethods in Enzymology
Volume518
ISSN (Print)00766879
ISSN (Electronic)15577988

Fingerprint

Fluorescence
Cells
Fluorescence Spectrometry
Spectroscopy
Sampling
Signal-To-Noise Ratio
Molecular interactions
Photons
Luminance
Experiments

Keywords

  • Brightness analysis
  • Cumulant analysis
  • Fluorescence correlation spectroscopy
  • Fluorescence fluctuation spectroscopy
  • Live cell
  • Photon counting histogram

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Wu, B., Singer, R. H., & Mueller, J. D. (2013). Time-integrated fluorescence cumulant analysis and its application in living cells. In Methods in Enzymology (Vol. 518, pp. 99-119). (Methods in Enzymology; Vol. 518). https://doi.org/10.1016/B978-0-12-388422-0.00005-4

Time-integrated fluorescence cumulant analysis and its application in living cells. / Wu, Bin; Singer, Robert H.; Mueller, Joachim D.

Methods in Enzymology. Vol. 518 2013. p. 99-119 (Methods in Enzymology; Vol. 518).

Research output: Chapter in Book/Report/Conference proceedingChapter

Wu, B, Singer, RH & Mueller, JD 2013, Time-integrated fluorescence cumulant analysis and its application in living cells. in Methods in Enzymology. vol. 518, Methods in Enzymology, vol. 518, pp. 99-119. https://doi.org/10.1016/B978-0-12-388422-0.00005-4
Wu B, Singer RH, Mueller JD. Time-integrated fluorescence cumulant analysis and its application in living cells. In Methods in Enzymology. Vol. 518. 2013. p. 99-119. (Methods in Enzymology). https://doi.org/10.1016/B978-0-12-388422-0.00005-4
Wu, Bin ; Singer, Robert H. ; Mueller, Joachim D. / Time-integrated fluorescence cumulant analysis and its application in living cells. Methods in Enzymology. Vol. 518 2013. pp. 99-119 (Methods in Enzymology).
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