TIR fluorescence screening of cell membranes

Thomas Bruns; Wolfgang S.L. Strauss; Reinhard Sailer; Michael Wagner; Herbert Schneckenburger

doi:10.1117/12.590199

29 March 2005 TIR fluorescence screening of cell membranes

Thomas Bruns, Wolfgang S.L. Strauss, Reinhard Sailer, Michael Wagner, Herbert Schneckenburger

Proceedings Volume 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III; (2005) https://doi.org/10.1117/12.590199
Event: SPIE BiOS, 2005, San Jose, CA, United States

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.

Citation Download Citation

Thomas Bruns, Wolfgang S.L. Strauss, Reinhard Sailer, Michael Wagner, and Herbert Schneckenburger "TIR fluorescence screening of cell membranes", Proc. SPIE 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, (29 March 2005); https://doi.org/10.1117/12.590199

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