TRAINING & CAREERS
2017 BESIP Project
Laboratory of Molecular Biophysics, Optical Spectroscopy Section
Jay R. Knutson, Ph.D., Chief, OSS
Molly Molloy, Ph.D.
Alessio Andreoni, Ph.D.
Laboratory and Project Description
We develop and exploit the techniques of ultrafast laser microscopy. We develop state-of-the-art instruments that help us determine the local flexibility, size, and structure of complexes both in solution and inside living cells. We have recently developed new ‘superresolution’ methods to observe <100nm resolution images in fluorescence microscopy - using both STED and our own novel “STAQ” dyes (requiring much less STED power). For years, we have been developing 2-photon, lifetime and FCS microscopy methods to extend a detailed spectroscopic approach to the cellular level. We have, for example, mapped the free/bound ratio of NADH in cardiac mitochondria (2p lifetime) and quantified the binding of HIV nef to receptor CD4 by monitoring crosscorrelation in translation (FCS). We have collaborated with LCE in the creation of new CARS and “Total Emission” systems along with new optimization (eg., genetic) algorithms for data analysis. Depending on their interests, summer interns will all be introduced to fluorescence instruments and image/data acquisition plus:
- Help exploit superresolved acquisition of cell images in novel microscopes, using transfected versions of the STAQ architecture.
- Help test and exploit a novel fluorescence chimera that senses ischemic [O2] levels.
- Help develop computer software to “globally” analyze either features and flexibility of proteins and DNA, or micro/nanoscopic image sets in cells.
- Exploit FLIM (Fluorescence Lifetime Microscopy) to push STED/STAQ resolution below 400A.
Feel free to call for more information.