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Proceedings Paper

TIR fluorescence screening of cell membranes
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Paper Abstract

A novel setup for fluorescence measurements of surfaces of biological samples, in particular cell membranes, is described. The method is based on multiple total internal reflection (TIR) of a laser beam on the surface of a multi-well plate, such that 96 individual samples are excited simultaneously. Main prerequisites are an appropriate thickness and high transmission of the glass bottom, a non-cytotoxic adhesive, and appropriate glass rods for TIR illumination. Fluorescence from cell surface is detected simultaneously using an integrating CCD camera and appropriate optical filters. For validation of the system, transfected cells expressing a fluorescent membrane protein are used. In addition, intracellular translocation of green fluorescent protein kinase c upon stimulation is examined. The method appears well suitable for high throughput screening (HTS), since neither washing of the samples nor any re-adjustment of the equipment after changing of individual plates are necessary.

Paper Details

Date Published: 29 March 2005
PDF: 6 pages
Proc. SPIE 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, (29 March 2005); doi: 10.1117/12.590199
Show Author Affiliations
Thomas Bruns, Hochschule Aalen (Germany)
Wolfgang S.L. Strauss, Univ. Ulm (Germany)
Reinhard Sailer, Univ. Ulm (Germany)
Michael Wagner, Hochschule Aalen (Germany)
Herbert Schneckenburger, Hochschule Aalen (Germany)
Univ. Ulm (Germany)

Published in SPIE Proceedings Vol. 5699:
Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III
Dan V. Nicolau; Dan V. Nicolau; Jörg Enderlein; Ramesh Raghavachari; Robert C. Leif; Daniel L. Farkas, Editor(s)

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