Washburn University Chemistry Club

Topic:Hyperspectral Fluorescence Imaging of Biological Systems

Speaker:Dr. David M. Haaland, Senior Scientist, Sandia National Laboratories
Adjunct Professor, University of New Mexico

Abstract:
We have developed a 3D hyperspectral confocal fluorescence microscope that can optically section live cells with submicron spatial resolution. For each voxel within the specimen, the microscope records the entire emission spectrum from 500 nm to 800 nm. When coupled with multivariate curve resolution (MCR), the new microscope can “discover,” quantify, and resolve multiple spatially and spectrally overlapped emission components, thereby greatly increasing the number of fluorescent labels that can be monitored simultaneously. The design and operation of the microscope will be discussed and a detailed description of the multivariate methods used to analyze the hyperspectral images will be presented. The utility of the new approach will be demonstrated with a number of hyperspectral images from biological samples. For example, we have imaged live Synechocystis cells with the new microscope. Synechocystis is a cyanobacterium, a class of organisms responsible for a large fraction of carbon sequestration from the atmosphere. In these experiments, cells from wild type cells and from mutants lacking a photosystem or a step in chlorophyll biosynthesis were imaged to better understand relative concentrations and spatial distributions of photosynthetic pigments in these bacteria. Six autofluorescence species were resolved in the cells, and for the first time the heterogeneity of pigment distribution within and between cells was observed. Hyperspectral images obtained from other biological systems including corn plant leaves, HeLa cells interacting with cytokines, multiple fluorescent proteins in macrophage cells, and Rat Basophilic Leukemia cells labeled with multiple-color quantum dots will also be presented. The new microscope and associated multivariate analyses constitute an enabling new technology for cell imaging and for understanding a variety of molecular and physical processes occurring in live cells.

Sandia is a multi-program laboratory operated by Sandia Corporation, a Lockheed Martin Company, for the United States Department of Energy under Contract DE-ACO4-94AL85000. A portion of this work was supported by the Microscale Immune Studies Laboratory Grand Challenge LDRD.

When: Thursday, October 23, 2008
Where: Buehler 415, University of Tennessee Main Campus.

All students and faculty are invited to attend.

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