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Proceedings Paper

Short-pulsed diode lasers as an excitation source for time-resolved fluorescence applications and confocal laser scanning microscopy in PDT
Author(s): Matthias Kress; Thomas H. Meier; Tarek Abd Allah El-Tayeb; Ralf Kemkemer; Rudolf W. Steiner; Angelika C. Rueck
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Paper Abstract

This article describes a setup for subcellular time-resolved fluorescence spectroscopy and fluorescence lifetime measurements using a confocal laser scanning microscope in combination with a short pulsed diode laser for fluorescence excitation and specimen illumination. The diode laser emits pulses at 398 nm wavelength with 70 ps full width at half maximum (FWHM) duration. The diode laser can be run at a pulse repetition rate of 40 MHz down to single shot mode. For time resolved spectroscopy a spectrometer setup consisting of an Czerny Turner spectrometer and a MCP-gated and -intensified CCD camera was used. Subcellular fluorescence lifetime measurements were achieved using a time-correlated single photon counting (TCSPC) module instead of the spectrometer setup. The capability of the short pulsed diode laser for fluorescence imaging, fluorescence lifetime measurements and time-resolved spectroscopy in combination with laser scanning microscopy is demonstrated by fluorescence analysis of several photosensitizers on a single cell level.

Paper Details

Date Published: 2 November 2001
PDF: 6 pages
Proc. SPIE 4431, Photon Migration, Optical Coherence Tomography, and Microscopy, (2 November 2001); doi: 10.1117/12.447408
Show Author Affiliations
Matthias Kress, Univ. Ulm (Germany)
Thomas H. Meier, Univ. Ulm (Germany)
Tarek Abd Allah El-Tayeb, Cairo Univ. (Egypt)
Ralf Kemkemer, Univ. Ulm (Germany)
Rudolf W. Steiner, Univ. Ulm (Germany)
Angelika C. Rueck, Univ. Ulm (Germany)

Published in SPIE Proceedings Vol. 4431:
Photon Migration, Optical Coherence Tomography, and Microscopy
Stefan Andersson-Engels; Michael F. Kaschke, Editor(s)

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