Development of widefield time-resolved fluorescence anisotropy imaging (TR-FAIM) using a single photon avalanche diode (SPAD) array

Louis Obeid Mogridge*, Jakub Nedbal, Shirwa Awale, Jess Tallis, Emily J. Johnston, Francesco Mattioli Della Rocca, Susan J. Rosser, Robert K. Henderson, Klaus Suhling

*Corresponding author for this work

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

Abstract / Description of output

The time-resolved fluorescence anisotropy of a biological sample can reveal rotational dynamics and structure of a fluorescent probe’s local environment. Here, methods are presented towards developing widefield time-resolved fluorescence anisotropy imaging (TR-FAIM) using a single photon avalanche diode (SPAD) array (QuantICAM). Such detector allows for simultaneous time-correlated single photon counting (TCSPC) measurements in each pixel with single-photon sensitivity and picosecond time resolution. Our method integrates the SPAD array with a widefield microscope, and automated rotating polarizers in the excitation and emission pathways to demonstrate TR-FAIM. We have shown the robustness of the method through spectroscopic measurements of the fluorophore PM546, and we have demonstrated the usefulness of simultaneous FLIM and TR-FAIM for studying properties of plasma membranes in live yeast cells.
Original languageEnglish
Title of host publicationProceedings Volume 12847
Subtitle of host publicationMultiphoton Microscopy in the Biomedical Sciences XXIV
EditorsAmmasi Periasamy, Peter T. So
Place of PublicationBellingham
PublisherSPIE
Chapter0J
Number of pages19
Volume12847
ISBN (Electronic)9781510669543
ISBN (Print)9781510669536
DOIs
Publication statusPublished - 12 Mar 2024
EventSPIE BiOS, 2024, San Francisco, California, United States - The Moscone Center, 747 Howard St, San Francisco, CA 94103, San Francisco, United States
Duration: 28 Jan 202430 Jan 2024
https://spie.org/conferences-and-exhibitions/photonics-west/exhibitions/bios-expo?SSO=1

Conference

ConferenceSPIE BiOS, 2024, San Francisco, California, United States
Country/TerritoryUnited States
CitySan Francisco
Period28/01/2430/01/24
Internet address

Keywords / Materials (for Non-textual outputs)

  • Time-resolved fluorescence anisotropy imaging (TR-FAIM
  • single-photon avalanche diode (SPAD) array
  • time-correlated single photon counting (TCSPC)
  • , fluorescence lifetime imaging microscopy (FLIM)
  • live-cell imaging
  • microviscosity
  • intracellular diffusion

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