Abstract

A novel setup for selective fluorescence screening of surfaces from vital 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. Fluorescence from cell surfaces, in particular plasma membranes, is measured simultaneously using an integrating CCD camera and appropriate optical filters. For validation of the method, fluorescent membrane markers as well as transfected human breast cancer-cells expressing a fluorescent membrane protein were used. In addition, intracellular translocation of green fluorescent protein kinase c was examined upon stimulation. 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. In view of an extension towards high content screening (HCS), additional data on fluorescence lifetime and fluorescence polarisation are presented.

© 2005 Optical Society of America