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A MICROSPECTROFLUOROMETER WITH EPI-ILLUMINATION OPERATED UNDER COMPUTER CONTROL
Author(s) -
J. S. Ploem,
J. A. DE STERKE,
J. Bonnet,
H. Wasmund
Publication year - 1974
Publication title -
journal of histochemistry and cytochemistry
Language(s) - English
Resource type - Journals
eISSN - 1551-5044
pISSN - 0022-1554
DOI - 10.1177/22.7.668
Subject(s) - monochromator , optics , fluorescence , intensity (physics) , line (geometry) , materials science , physics , mathematics , wavelength , geometry
Until recently the recording of fully corrected excitation and fluorescence spectra from fluorophores in microscopic preparations has been a time-consuming procedure. A new type of microspectrofluorometer with epi-illumination has been developed. This instrument is operated on line with a digital computer permitting automatic operation of monochromator motors, electrical shutters and correction of spectra. The electrical shutters are used for a rapid change between low intensity dia- and strong epi-illumination. Exposure to epi-illumination is limited to the short period of recording the spectra. A new optical system permits centering of structures to be measured, using the binocular head for viewing the illuminated image of the monochromator slit and the fluorescent image excited with dia-illumination simultaneously. New objectives and eyepieces with a high light collecting power permit the visualization of weak fluorescence obtained with low intensity dia-illumination.

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