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AKB: Biologische Physik

AKB 200: Poster Session II

AKB 200.65: Poster

Tuesday, March 8, 2005, 17:00–19:00, Poster TU C

A grating based detection platform for multi-color fluorescence correlation spectroscopy — •Markus Burkhardt, Katrin G. Heinze, and Petra Schwille — Biotec/TU Dresden, Tatzberg 47-51, 01307 Dresden, Germany

Fluorescence Correlation Spectroscopy (FCS) is based on time dependent fluorescence intensity fluctuations of labeled biomolecules as they enter and leave a diffraction-limited optical detection volume via Brownian diffusion. From these fluctuations, concentrations, diffusion and binding coefficients are easily obtained. Simultaneous monitoring of biomolecular species, labeled with spectrally distinct fluorophores, has proven to reveal inter- and intramolecular mechanisms both in vitro and in vivo.

We have developed a continuously tunable, filterless multi-color detection unit for FCS. Our tailored platform, using a grating instead of the classical dichroic mirror cascade, allows for accommodation of up to 15 detection channels covering the entire visible spectral range. As a proof of principle, we successfully demonstrate simultaneous FCS of four distinct fluorescent quantum dots being mixed in aqueous solution. Grating-based detection allows for spectral high-resolution FCS and is a feasible tool for quantitative investigation of complex biomolecular dynamics on a single molecule level.