siFLIM: single-image frequency-domain FLIM provides fast and photon-efficient lifetime data

Nat Methods. 2016 Jun;13(6):501-4. doi: 10.1038/nmeth.3836. Epub 2016 Apr 18.


We developed single-image fluorescence lifetime imaging microscopy (siFLIM), a method for acquiring quantitative lifetime images from a single exposure. siFLIM takes advantage of a new generation of dedicated cameras that simultaneously record two 180°-phase-shifted images, and it allows for video-rate lifetime imaging with minimal phototoxicity and bleaching. siFLIM is also inherently immune to artifacts stemming from rapid cellular movements and signal transients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium / metabolism
  • Calibration
  • Fluorescent Dyes / chemistry
  • HeLa Cells
  • Histamine / pharmacology
  • Humans
  • Image Enhancement / methods*
  • Image Interpretation, Computer-Assisted / methods*
  • Microscopy, Fluorescence / methods*
  • Photons
  • Time-Lapse Imaging / methods*


  • Fluorescent Dyes
  • Histamine
  • Calcium