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Cited by 613Advanced Fluorescence Microscopy TechniquesCell Image Analysis TechniquesAdvanced Measurement and Detection Methods

Accuracy and precision in quantitative fluorescence microscopy

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Waters, Jennifer C.·2009·Journal of Cell Biology
DOI 10.1083/jcb.200903097PMC PMC2712964
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Abstract

The light microscope has long been used to document the localization of fluorescent molecules in cell biology research. With advances in digital cameras and the discovery and development of genetically encoded fluorophores, there has been a huge increase in the use of fluorescence microscopy to quantify spatial and temporal measurements of fluorescent molecules in biological specimens. Whether simply comparing the relative intensities of two fluorescent specimens, or using advanced techniques like Förster resonance energy transfer (FRET) or fluorescence recovery after photobleaching (FRAP), quantitation of fluorescence requires a thorough understanding of the limitations of and proper use of the different components of the imaging system. Here, I focus on the parameters of digital image acquisition that affect the accuracy and precision of quantitative fluorescence microscopy measurements.

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Cited by

Sorted by citations

Authors
Waters, J. C.
Affiliations
Harvard University (US)
Corresponding author
Waters, Jennifer C.
ORCID
Waters, Jennifer C.
Venue
Journal of Cell Biology
Publisher
Rockefeller University Press
Published
2009-06-29
Volume
185
Issue
7
Pages
1135-1148
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
DOI
10.1083/jcb.200903097
PMC
PMC2712964
OpenAlex
W2141201649
Cited by
613
References
1
Citations by year
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FWCI
9.31
Type
journal article
Language
en
Open access
Licence
cc-by-nc-sa
Locations
  • The Journal of Cell Biology (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
  • Physical Sciences › Engineering › Electrical and Electronic Engineering › Advanced Measurement and Detection Methods
Keywords
quantitative fluorescence microscopy, measurement accuracy, measurement precision, digital image acquisition, image acquisition parameters, fluorescence intensity measurement
MeSH terms
Fluorescent Dyes, Image Interpretation, Computer-Assisted, Light, Microscopy, Fluorescence, Sensitivity and Specificity

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