Application of a novel 8x8 PMT array detector to light microscopy

Kenneth R. Spring Jr.; Olga N. Kovbasnjuk; Carter C. Gibson; Peter M. Bungay

doi:10.1117/12.310554

9 June 1998 Application of a novel 8x8 PMT array detector to light microscopy

Kenneth R. Spring Jr., Olga N. Kovbasnjuk, Carter C. Gibson, Peter M. Bungay

Proceedings Volume 3261, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing V; (1998) https://doi.org/10.1117/12.310554
Event: BiOS '98 International Biomedical Optics Symposium, 1998, San Jose, CA, United States

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.

Citation Download Citation

Kenneth R. Spring Jr., Olga N. Kovbasnjuk, Carter C. Gibson, and Peter M. Bungay "Application of a novel 8x8 PMT array detector to light microscopy", Proc. SPIE 3261, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing V, (9 June 1998); https://doi.org/10.1117/12.310554

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