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Proceedings Paper

Application of a novel 8x8 PMT array detector to light microscopy
Author(s): Kenneth R. Spring; Olga N. Kovbasnjuk; Carter C. Gibson; Peter M. Bungay
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Paper Abstract

A multi-anode photomultiplier tube (PMT) attached to a light microscope was used to measure the low-light-level signals from fluorescent dyes trapped in the extracellular space between living, cultured kidney epithelial cells. The detection assembly utilized photon counting of all 64 channels to record the fluorescence produced after the photoactivation of caged fluorophores of different molecular weight and charge. Photoactivation was accomplished by a brief pulse of light at 355 nm forma frequency-tripled Nd:YAG laser. Fluorescence of the uncaged fluorophores was excited by the output of an argon laser equipped with an acousto-optical tunable filter for control of wavelength and power. Diffusion coefficients for the fluorescent indicators in the extracellular spaces were calculated from the spatial and temporal decay of the fluorescence after uncaging of the dye in a small region micrometers -diameter microscope field. The system magnification was adjusted so that each 2.5-mm-square PMT channel corresponded to a 12.5-micrometers square region of the microscope field. The spatial decay of the fluorescence was obtained by sampling multiple adjacent PMT channels, while the temporal decay was determined from the PMT channel encompassing the uncaging site.

Paper Details

Date Published: 9 June 1998
PDF: 4 pages
Proc. SPIE 3261, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing V, (9 June 1998); doi: 10.1117/12.310554
Show Author Affiliations
Kenneth R. Spring, National Institutes of Health (United States)
Olga N. Kovbasnjuk, National Institutes of Health (United States)
Carter C. Gibson, National Institutes of Health (United States)
Peter M. Bungay, National Institutes of Health (United States)

Published in SPIE Proceedings Vol. 3261:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing V
Thomas Taiwei Lu; Carol J. Cogswell; Jeremy M. Lerner; Jose-Angel Conchello; Jeremy M. Lerner; Thomas Taiwei Lu; Tony Wilson, Editor(s)

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