Stroboscopic fluorescence lifetime imaging

Holton, Mark D., Silvestre, Oscar, Errington, Rachel Jane ORCID: https://orcid.org/0000-0002-8016-4376, Smith, Paul James, Matthews, Daniel R., Rees, Paul and Summers, Huw D. 2009. Stroboscopic fluorescence lifetime imaging. Optics Express 17 (7) , pp. 5205-5216. 10.1364/OE.17.005205

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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 intra-cellular 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.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Subjects:	R Medicine > R Medicine (General)
Additional Information:	Pdf uploaded in accordance with publisher's policy at http://www.sherpa.ac.uk/romeo/issn/1094-4087/ and http://www.opticsinfobase.org/submit/review/copyright_permissions.cfm#posting (accessed 28/02/2014).
Publisher:	Optical Society of America
ISSN:	1094-4087
Date of First Compliant Deposit:	30 March 2016
Last Modified:	09 May 2023 15:10
URI:	https://orca.cardiff.ac.uk/id/eprint/37108

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