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Electron Microscopic Demonstration of the Photoconversion of a Fluorescent Label in Mixed Neuroglial Cultures

Published online by Cambridge University Press:  02 July 2020

Kelly Ann Randall ,
Cristian L. Achim ,
Clayton A. Wiley  and
Linda M. Dallasta
Kelly Ann Randall
Affiliation:
University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213
Cristian L. Achim
Affiliation:
University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213
Clayton A. Wiley
Affiliation:
University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213
Linda M. Dallasta
Affiliation:
University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213
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Extract

Endothelial cells (EC) possess receptors for and metabolize low-density lipoprotein (LDL); acetylation of LDL promotes EC metabolism and the intracellular deposition of its cholesteryl ester component. In turn, acetylated LDL linked to the fluorescent dye, Dil (Dil-Ac-LDL), has been used as a marker for endothelial cells, including cerebral microvascular endothelial cells (CEC). Due to the low electron density of fluorescent dyes, however, detection of this marker is not possible ultrastructurally. We demonstrate here the photo-conversion of the chromogen, 3,3'-diaminobenzidine tetrahydrochloride (DAB), into a stable, electron-dense reaction product in subcultured human fetal CEC containing the fluorescent marker, Dil-Ac-LDL, thus promoting identification of these cells via light and electron microscopy. This method, in turn, may be useful in identifying and tracking human fetal CEC grafted into SCID mice as a model for blood-brain barrier angiogenesis.

Isolated human fetal CEC were subcultured and labeled with Dil-Ac-LDL (10 ug/ml).

Type
Technologists’ Forum: Special Topics and Symposium
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 367 - 368
Copyright
Copyright © Microscopy Society of America 1997

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