Stroboscopic fluorescence lifetime imaging

Mark D. Holton, Oscar R. Silvestre, Rachel J. Errington, Paul J. Smith, Daniel R. Matthews, Paul Rees, Huw D. Summers

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

We report a fluorescence lifetime imaging technique that uses the time integrated response to a periodic optical excitation, eliminating the need for time resolution in detection. A Dirac pulse train of variable period is used to probe the frequency response of the total fluorescence per pulse leading to a frequency roll-off that is dependent on the relaxation rate of the fluorophores. The technique is validated by demonstrating wide-field, realtime, lifetime imaging of the endocytosis of inorganic quantum dots by a cancer cell line. Surface charging of the dots in the intracellular environment produces a switch in the fluorescence lifetime from ∼ 40 ns to < 10 ns. A temporal resolution of half the excitation period is possible which in this instance is 15 ns. This stroboscopic technique offers lifetime based imaging at video rates with standard CCD cameras and has application in probing millisecond cell dynamics and in high throughput imaging assays.

Original languageEnglish (US)
Pages (from-to)5205-5216
Number of pages12
JournalOptics Express
Volume17
Issue number7
DOIs
StatePublished - Mar 30 2009

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics

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