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 Citation (Scopus)

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
Volume2412
ISBN (Print)0819417599
StatePublished - 1995
Externally publishedYes
EventThree-Dimensional Microscopy: Image Acquisition and Processing II - San Jose, CA, USA
Duration: Feb 9 1995Feb 10 1995

Other

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

Fingerprint

Confocal microscopy
Laser pulses
Fluorescence
microscopy
Imaging techniques
life (durability)
fluorescence
signal detection
Signal detection
pulses
Charge coupled devices
excitation
dynamic range
charge coupled devices
pulse duration
Molecules
probes
electronics
lasers
molecules

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

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 (Vol. 2412, pp. 115-123). Society of Photo-Optical Instrumentation Engineers.

New method for fluorescence lifetime imaging in bilateral-confocal microscopy by double-pulse excitation. / Brakenhoff, G. J.; Mueller, M.; Ghauharali, R. I.; Visscher, Koen.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / Tony Wilson; Carol J. Cogswell. Vol. 2412 Society of Photo-Optical Instrumentation Engineers, 1995. p. 115-123.

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

Brakenhoff, GJ, Mueller, M, Ghauharali, RI & Visscher, K 1995, New method for fluorescence lifetime imaging in bilateral-confocal microscopy by double-pulse excitation. in T Wilson & CJ Cogswell (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 2412, Society of Photo-Optical Instrumentation Engineers, pp. 115-123, Three-Dimensional Microscopy: Image Acquisition and Processing II, San Jose, CA, USA, 2/9/95.
Brakenhoff GJ, Mueller M, Ghauharali RI, Visscher K. New method for fluorescence lifetime imaging in bilateral-confocal microscopy by double-pulse excitation. In Wilson T, Cogswell CJ, editors, Proceedings of SPIE - The International Society for Optical Engineering. Vol. 2412. Society of Photo-Optical Instrumentation Engineers. 1995. p. 115-123
Brakenhoff, G. J. ; Mueller, M. ; Ghauharali, R. I. ; Visscher, Koen. / New method for fluorescence lifetime imaging in bilateral-confocal microscopy by double-pulse excitation. Proceedings of SPIE - The International Society for Optical Engineering. editor / Tony Wilson ; Carol J. Cogswell. Vol. 2412 Society of Photo-Optical Instrumentation Engineers, 1995. pp. 115-123
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