New method for fluorescence lifetime imaging in bilateral-confocal microscopy by double-pulse excitation

G. J. Brakenhoff, M. Mueller, R. I. Ghauharali, Koen Visscher

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

1 Scopus citations

Abstract

A new technique for the measurement of fluorescence lifetimes relies on the (near steady state) excitation with short optical pulses. The novel technique has the potentiality to provide high time resolution - since it relies on the laser pulse duration, rather than on electronic gating techniques - and permits, in combination with bilateral confocal microscopy and the use of a (cooled) CCD, sensitive signal detection over a large dynamic range. Combined with confocal microscopy it enables the spatial determination of the fluorescence lifetimes, the value of which is influenced by the local environment of fluorescent probe molecules in biological samples. The principle of the technique is discussed within a theoretical framework taking into account various secondary effects.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsTony Wilson, Carol J. Cogswell
PublisherSociety of Photo-Optical Instrumentation Engineers
Pages115-123
Number of pages9
ISBN (Print)0819417599
StatePublished - Jan 1 1995
Externally publishedYes
EventThree-Dimensional Microscopy: Image Acquisition and Processing II - San Jose, CA, USA
Duration: Feb 9 1995Feb 10 1995

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume2412
ISSN (Print)0277-786X

Other

OtherThree-Dimensional Microscopy: Image Acquisition and Processing II
CitySan Jose, CA, USA
Period2/9/952/10/95

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ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

Cite this

Brakenhoff, G. J., Mueller, M., Ghauharali, R. I., & Visscher, K. (1995). New method for fluorescence lifetime imaging in bilateral-confocal microscopy by double-pulse excitation. In T. Wilson, & C. J. Cogswell (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (pp. 115-123). (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 2412). Society of Photo-Optical Instrumentation Engineers.