Overview of Fluorescence Lifetime Measurements in Flow Cytometry

The focus of this chapter is time-resolved flow cytometry, which is broadly defined as the ability to measure the timing of fluorescence decay from excited fluorophores that pass through cytometers or high-throughput cell counting and cell sorting instruments. We focus on this subject for two main r...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1678; p. 421
Main Authors Houston, Jessica P, Yang, Zhihua, Sambrano, Jesse, Li, Wenyan, Nichani, Kapil, Vacca, Giacomo
Format Journal Article
LanguageEnglish
Published United States 01.01.2018
Subjects
Flow Cytometry - instrumentation
Flow Cytometry - methods
Fluorescence
Fluorescence Resonance Energy Transfer
Fluorescent Dyes
Kinetics
Luminescent Proteins
Time Factors
Workflow
FRET
Fluorescence lifetime
Time-domain
Frequency-domain
Time-resolved flow cytometry
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Summary:The focus of this chapter is time-resolved flow cytometry, which is broadly defined as the ability to measure the timing of fluorescence decay from excited fluorophores that pass through cytometers or high-throughput cell counting and cell sorting instruments. We focus on this subject for two main reasons: first, to discuss the nuances of hardware and software modifications needed for these measurements because currently, there are no widespread time-resolved cytometers nor a one-size-fits-all approach; and second, to summarize the application space for fluorescence lifetime-based cell counting/sorting owing to the recent increase in the number of investigators interested in this approach. Overall, this chapter is structured into three sections: (1) theory of fluorescence decay kinetics, (2) modern time-resolved flow cytometry systems, and (3) cell counting and sorting applications. These commentaries are followed by conclusions and discussion about new directions and opportunities for fluorescence lifetime measurements in flow cytometry.
ISSN:1940-6029
DOI:10.1007/978-1-4939-7346-0_18