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Greece : Picosecond fluorescence lifetime imaging microscopy - application to single molecule dynamics.

Research objectives and content: Objectives: To set up a time- and space-correlated single photon counti (TSCSPC) spectroscopy system by using (a) the pico-second pulsed laser system of the host and (b) the TSCSPC detection system of the guest. There will be two spectroscopy systems: (a) delay-line (DL) microchannel plate (MCP) photomultiplier (PMT) attached to a polychromator and (b) quadrant-anode (QA) MCP-PMT attached to a low-background microscope, where DL- and QA-MCP-PMT are linear and imaging detectors of 10 ps time resolution. respectively. Content: To perform time-resolved spectroscopy of individual dye molecules, relevant in fundamental spectroscopy, biotechnology, and medical research. The kinetics of individual molecules may differ significantly from the average values of an ensemble. Individual kinetics of probe molecules will be of considerable interest in structural molecular biology. The antifungal agent nystatin, for instance, is an intrinsically fluorescent molecule that stresses membranes. The population of the antibiotics is believed to be divided in two sub-populations having different locations and fluorescence lifetimes, probably corresponding to different aggregation states Ensemble average measurements prevent any detailed structural inforrnation of such an heterogeneous system. The use of a sensitive time-resolved spectroscopy will solve a long time open question: What is the role of different aggregation states in the biochemical mode of action of nystatin? The use of TSCSPC spectroscopy would also be very welcome to study the binding of the hemagglutinin (HA) protein from Influenza virus to sialic acid residues.

country :Greece

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Publication:Mena Report
Date:Feb 8, 2013
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