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Confocal redox fluorescence microscopy for the evaluation of corneal hypoxia
Conference proceeding

Confocal redox fluorescence microscopy for the evaluation of corneal hypoxia

Barry R Masters, Andres Kriete and Joerg Kukulies
Proceedings of SPIE, v 1431(1)
01 May 1991
DOI: https://doi.org/10.1117/12.44192

Abstract

A Zeiss laser scanning microscope was fitted with a high powered Argon ion laser (10 W) which provided wavelengths in the following regions: 364 nm (multiline), 488 nm and 514 nm. A Zeiss water object of 40X, NA. 0.6, corrected for the UV was used to measure the fluorescence from optical sections of a freshly enucleated rabbit eye. The resolution in the transverse direction was about 0.5 micron and the range resolution was about 0.7 micron at 366 nm wavelengths. The confocal microscope was used in both the reflected mode and the confocal mode to image the endothelial cells of the enucleated eye. Reflected light images were obtained at all wavelengths from the argon laser, and also from the HeNe laser line at 633 nm was used to image the cells in reflected light. The same fields of cells were imaged in fluorescence light. The wavelengths of excitation of 366 nm for the excitation and 400-500 for the emission were used to image the pyridine nucleotides. The reduced pyridine nucleotides are suitable chromophores for the evaluation of cellular hypoxia in the living eye. This paper demonstrated the feasibility of two dimensional fluorescent imaging of the reduced pyridine nucleotides in the corneal endothelial cells. The confocal image was made through 400 microns of corneal tissue.

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Web of Science research areas
Medicine, Research & Experimental
Spectroscopy
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