Manchester Centre for Biophysics and Catalysis (MCBC)

Single molecule approaches

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MCBC is developing novel single molecule methods to study the dynamics of macromolecular systems. We have a fluorescence set-up based on confocal microscopy with pulsed laser excitation and simultaneous photon-counting detection in four channels (two-colour for both parallel and perpendicular polarization). Data are recorded either as a single molecule traverses the focal volume (repeated for thousands of individual single-molecule events), or by looking at time traces of an immobilized molecule. This includes total internal reflected fluorescence (TIRF) for high-throughput measurements, which are particularly important when developing surface and immobilization chemistry. TIRF will be implemented in a multi-parameter configuration (analogous to confocal above), by splitting the fluorescence onto different parts of a CCD camera (e.g. two colour and two polarization). A variety of pulsed and CW laser sources allow access to the wide range of fluorescent molecules. Single molecule instrumentation is located in both the Photon Science Institute and the Manchester Interdisciplinary Biocentre.

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