SINGLE PHOTON COUNTING FLUORESCENCE & IMAGING APPARATUS

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The time-correlated single photon counting (TCSPC) fluorescence apparatus performs as a state-of-the art spectroscopic instrument. In accord with the goal of developing new technologies out of existing instrumentation we have developed a time resolved confocal optical spectrometer which revolutionizes the study of molecular dynamic and photonic processes by incorporating the coupling the spatial resolution with state-of-the-art ultrafast spectroscopic methods. Our instrument is designed to acquire single location or scanned images of samples using three contrast mechanisms (force, intensity and excited state lifetime) in quickly converting configurations. Within the past year we have updated our confocal microscope to two channel operation which simultaneously allows detection of photons in two separate spectral regions. This allows us the capability of monitoring resonance fluorescence energy transfer between donor/acceptor pairs in many different systems; among those recently studies are tagged GCN-4 peptides and intracellular proteins in vivo. Other developments include the separation of the conventional TCSPC system from the confocal optical spectrometer. This separation in detection and electronics, which will facilitate maximum use of both instruments, is nearly complete. Only the updating and programming of a separate computer data acquisition center (with a faster processor) for the TCSPC remains to be completed.