FIP Seminar - Dr. Kevin Welsher "Real-time 3D microscopy methods"
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Wed, 09/21/2016 - 12:00 to 13:00
Dr. Kevin Welsher
Dr. Kevin Welsher, Asst. Prof., Duke University "Real-time 3D microscopy methods" - Single molecule spectroscopy elucidates the internal dynamics of immobilized molecules in vitro by collecting a continuous stream of emitted photons on a single photon counting detector, allowing continuous observation of the real-time molecular state, temporally limited only by the photon emission rate. In live cells, molecules rapidly diffuse out of the observation volume, precluding continuous observation. Moreover, in contrast to in vitro experiments, the molecule's cellular environment becomes critical to its dynamics. To enable continuous observation of molecular species in their live-cell context we developed 3D Multi-resolution Microscopy (3D-MM). 3D-MM utilizes real-time 3D single-particle tracking to "lock-on" to a single diffusing nanoparticle using optical feedback and a 3D piezo-electric stage, permitting continuous observation of its dynamics. The cellular context is collected via a two-photon laser scanning microscope, which constructs a 3D map of the nanoparticle's surroundings.