Fluorescence imaging and time-resolved spectroscopy of steroid using confocal synchrotron radiation microscopy

Hans C. Gerritsen; C. J. R. van der Oord; Yehudi K. Levine; Ian H. Munro; Gareth R. Jones; D. A. Shaw; Fokko F.G. Rommerts

doi:10.1117/12.182732

17 August 1994 Fluorescence imaging and time-resolved spectroscopy of steroid using confocal synchrotron radiation microscopy

Hans C. Gerritsen, C. J. R. van der Oord, Yehudi K. Levine, Ian H. Munro, Gareth R. Jones, D. A. Shaw, Fokko F.G. Rommerts

Author Affiliations +

Proceedings Volume 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV; (1994) https://doi.org/10.1117/12.182732
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

The Confocal Synchrotron Radiation Microscope at Daresbury was used in a study of the transport and distribution of the steroid Coumestrol in single Leydig cells. The broad spectrum of synchrotron radiation in combination with UV compatible microscope optics affords the extension of confocal microscopy from the visible to the UV region down to about 200 nm. Consequently fluorescent molecules with absorption bands in the UV can be imaged. In addition the pulsed nature of the light source allows us to perform time-resolved fluorescence spectroscopy experiments on microscopic volumes. Coumestrol is a naturally fluorescing plant steroid exhibiting estrogenic activity. In physiological environments it has an absorption peak in the UV at 340 nm and it emits around 440 nm. First results indicate that the Coumestrol transport through the cell membrane is diffusion limited. The weak fluorescence observed in the nuclei of the Leydig cells may be due to fluorescence quenching arising from the interaction of the Coumesterol with nuclear components. However, micro-volume time-resolved fluorescence spectroscopy experiments on cell nuclei have revealed the same decay behavior for Coumesterol in both the cytoplasm and nucleus of the cells.

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Hans C. Gerritsen, C. J. R. van der Oord, Yehudi K. Levine, Ian H. Munro, Gareth R. Jones, D. A. Shaw, and Fokko F.G. Rommerts "Fluorescence imaging and time-resolved spectroscopy of steroid using confocal synchrotron radiation microscopy", Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); https://doi.org/10.1117/12.182732

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