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Cited by 55Advanced Fluorescence Microscopy TechniquesPhotochemistry and Electron Transfer StudiesCell Image Analysis Techniques

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Fluorescence Lifetime Imaging Microscopy for Quantitative Biological Imaging

Chen, Leng-Chun, Lloyd, William R., Chang, Ching-Wei, Sud, Dhruv, Mycek, Mary-Ann·2013·Methods in Cell Biology
DOI 10.1016/b978-0-12-407761-4.00020-8
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Abstract

Fluorescence lifetime imaging microscopy (FLIM) is a method for measuring fluorophore lifetimes with microscopic spatial resolution, providing a useful tool for cell biologists to detect, visualize, and investigate structure and function of biological systems. In this chapter, we begin by introducing the basic theory of fluorescence lifetime, including the characteristics of fluorophore decay, followed by a discussion of factors affecting fluorescence lifetimes and the potential advantages of fluorescence lifetime as a source of image contrast. Experimental methods for creating lifetime maps, including both time- and frequency-domain experimental approaches, are then introduced. Then, FLIM data analysis methods are discussed, including rapid lifetime determination, multiexponential fitting, Laguerre polynomial fitting, and phasor plot analysis. After, data analysis methods are introduced that improve lifetime precision of FLIM maps based upon optimal virtual gating and total variation denoising. The chapter concludes by highlighting several recent FLIM applications for quantitative biological imaging, including Förster resonance energy transfer-FLIM, fluorescence correlation spectroscopy-FLIM, multispectral-FLIM, and multiphoton-FLIM.

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  • Photoacoustic and fluorescence lifetime imaging of diatoms

    Cvjetinovic, J., Salimon, A. I., Novoselova, M. V., Sapozhnikov, P. V., Shirshin, E. A., Yashchenok, A. M., Kalinina, O. Y., Korsunsky, A. M., & Gorin, D. A. (2020) · Photoacoustics · doi:10.1016/j.pacs.2020.100171 · Cited by 18

    “As an intrinsic property of a fluorophore, the fluorescence lifetime depends on temperature, pH, concentration, polarity, the presence of fluorescence quenchers, and internal factors that are connected with fluorophore structure [18,19].”
    Background

See all 55

Metadata

Authors
Chen, L.-C., Lloyd, W. R., Chang, C.-W., Sud, D., & Mycek, M.-A.
Affiliations
University of Michigan (US)
ORCID
Venue
Methods in Cell Biology
Publisher
Elsevier
Published
2013
Volume
114
Pages
457-488
ISSN
0091-679X (ISSN-L)
DOI
10.1016/b978-0-12-407761-4.00020-8
OpenAlex
W44585643
Cited by
55
References
0
Citations by year
2026: 1 · 2025: 3 · 2024: 5 · 2023: 3 · 2022: 7 · 2021: 3 · 2020: 4 · 2019: 5 · 2018: 5 · 2017: 4 · 2016: 5 · 2015: 4 · 2014: 6
FWCI
6.38
Type
book chapter
Language
en
Open access
closed
Locations
  • Methods in cell biology (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health
  • National Institute of Dental and Craniofacial Research (R01-DE-01943)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Physical Sciences › Chemistry › Physical and Theoretical Chemistry › Photochemistry and Electron Transfer Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
Keywords
fluorescence lifetime imaging, fluorescence decay, phasor analysis, total variation denoising, fluorescence resonance energy transfer, fluorescence correlation spectroscopy, multispectral imaging, multiphoton imaging
MeSH terms
Algorithms, Data Interpretation, Statistical, Diffusion, Fluorescent Dyes, HeLa Cells, Humans, Kinetics, Microscopy, Fluorescence, Mitosis, Spectrometry, Fluorescence, Least-Squares Analysis, Protein Transport, Fluorescence Resonance Energy Transfer, Green Fluorescent Proteins, Plant Stomata, Single-Cell Analysis

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Chen, L.-C., Lloyd, W. R., Chang, C.-W., Sud, D., & Mycek, M.-A. (2013). Fluorescence Lifetime Imaging Microscopy for Quantitative Biological Imaging. Methods in Cell Biology. https://doi.org/10.1016/b978-0-12-407761-4.00020-8

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