Phasor approach for fluorescence anisotropy

Dongeun Kim, Wonsang Hwang, Dug Young Kim

Research output: Chapter in Book/Report/Conference proceedingConference contribution


Fluorescence anisotropy is a powerful tool for observing molecular rotational speed, which is widely applied for molecular conjugation measurements (quantitative assay, membrane tagging, and protein-protein interaction). Fluorescence anisotropy (r) can be obtained by subtracting horizontal polarized fluorescence (H) from vertical polarized fluorescence (V), and dividing by non-polarized fluorescence (F). Since F equals to V+2H, V and H can be thought of as weighted sum and subtraction of F and r·F, respectively. Using phasor approach, which is graphical plotting technique based on fluorescence lifetime imaging microscopy, V and H are located on internal and external dividing points of F and r·F. Considering those four phasor points (V, H, F, and r·F) lie on a straight line, one can easily guess the value of the r. We first introduced phasor plot to fluorescence anisotropy, and confirmed that this method dramatically simplifies fluorescence anisotropy analysis with graphical intuition.

Original languageEnglish
Title of host publicationSingle Molecule Spectroscopy and Superresolution Imaging XIV
EditorsIngo Gregor, Felix Koberling, Rainer Erdmann
ISBN (Electronic)9781510641358
Publication statusPublished - 2021
EventSingle Molecule Spectroscopy and Superresolution Imaging XIV 2021 - Virtual, Online, United States
Duration: 2021 Mar 62021 Mar 11

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
ISSN (Print)1605-7422


ConferenceSingle Molecule Spectroscopy and Superresolution Imaging XIV 2021
Country/TerritoryUnited States
CityVirtual, Online

Bibliographical note

Publisher Copyright:
© 2021.

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Biomaterials
  • Radiology Nuclear Medicine and imaging


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