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BioSpectroscopy Core Research Facility
| Description | Established as a research resource at The University of Montana (UM) in 2002
Directed by Dr. J.B. Alexander Ross, Professor of Chemistry and Biochemistry at UM
Associated with the Center for Biomolecular Structure and Dynamics (CBSD) and the Center for Structural and Functional Neuroscience (CSFN)
Mission
To facilitate the research programs of biomedical and chemical investigators by providing state-of-the-art spectroscopy and microscopy tools
The BioSpectroscopy Core Research Facility (CRF) integrates its research and service activities with other UM CRFs to provide multidisciplinary approaches and collaborations from within and without the University to address fundamental questions in biophysics, biochemistry and chemistry and applied problems in human health and environmental quality
Tools
The BioSpectroscopy Core enables analysis of the structure, function, and interactions of biological macromolecules in solution, in cells, on model membranes, and on other surfaces using steady-state and time-resolved fluorescence spectroscopies, single- and multi-photon fluorescence correlation spectroscopy (FCS), and fluorescence lifetime imaging microscopy (FLIM). The Core also has the following capabilities for microscopy and imaging: direct excitation fluorescence, total internal reflectance fluorescence (TIRF), time-resolved fluorescence anisotropy, fluorescence lifetime imaging (FLIM), fluorescence correlation spectroscopy (FCS), and single-molecule Förster resonance energy transfer (sm-FRET).
Structure
The BioSpectroscopy CRF is structured according to the six elements of a National Institutes of Health (NIH) Biomedical Technology Program (P41), supported by the National Center for Research Resources (NCRRi):
(1) Technology research and development (R&D)
(2) Collaborative CRF research activities As characteristic of NCRR Research Resources, the collaborations help stimulate R&D, and R&D provides new tools for investigators.
(3) Service to intramural and extramural investigators Provides access to specialized instrumentation, software and experimental techniques
(4) Training to investigators and their students
(5) Dissemination of information Papers and presentations, website (in development) with links to CSFN and CBSD and to the other associated CRFs
(6) Oversight by a scientific advisory committee Composed of the UM Vice-President for Research, the CSFN and CBSD Directors and an ad hoc panel of scientists with expertise in the activities emphasized in the BioSpectroscopy CRF
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| Specialty equipment/services | • Time-correlated, single-photon counting (TCSPC) fluorometer. The fully automated sample chamber, designed and constructed in collaboration with Quantum Northwest (Liberty Lake, WA), uses a modified “T-format” optical arrangement that permits the simultaneous collection of multiple decay curves. On one side, a standard polarizer is used to collect decays at any angle, including magic angle. On the other side, a beam-splitting polarizer is used to separate the vertical and horizontal components of the emission. There are separate emission trains for the three separate polarization components. Each emission train uses monochromators or filters followed by separate photomultiplier tubes. Each of the three emission detectors has dedicated processing electronics. The sample chamber can accommodate different sample holders, including standard temperature-controlled cuvettes, a high-pressure cell (up to 2.5 kbar), or a goniometer for crystal samples. • Nikon TE2000U inverted microscope for time-resolved fluorescence spectroscopy using single or multi-photon excitation. This microscope has four ports for observation, and has been configured for direct excitation fluorescence, total internal reflectance fluorescence (TIRF) as well as time-resolved fluorescence anisotropy experiments. • Olympus IX71 inverted microscope equipped with FluoView 300 confocal scan-head for multi- and single-photon excitation This microscope system is used for fluorescence lifetime imaging (FLIM), fluorescence correlation spectroscopy (FCS) and single-molecule Förster resonance energy transfer (sm-FRET) experiments. • Ultra-fast lasers (1) A Coherent laser system that consists of a 10 W diode laser (Verde) pumping a Ti:Sapphire laser (Mira), which can be mode-locked to provide fs- or ps-wide pulses of light, is used as a light source for the TCSPC fluorometer as well as for the Nikon and Olympus microscopes. The Ti:Sapphire is tunable to wavelengths between ~680 and ~1050 nm. With second and third harmonic generation, UV and visible light can be produced from ~260 to ~520 nm. The Ti:Sapphire laser also can be used to pump an optical parametric oscillator (OPO) to generate ~1040 to ~1300 nm light, which frequency doubled provides ~520 to ~650 nm light. Thus, this laser system can provide pulsed light from the UV into the visible, as the near IR, to provide either direct or two-photon excitation of most intrinsic or extrinsic probes used to study biological systems. (2) A second Coherent Ti:Sapphire fs-pulsed laser (Chameleon) is dedicated to two- and three-photon microscopy experiments. • Specialized analysis software PicoQuant FluoFit (reconvolution lifetime and time-resolved anisotropy), PicoQuant SymPhoTime (FCS, FLIM, FRET), OriginLab (general data fitting). |
| Web address(es) | BioSpectroscopy Core Research Facility |
| Location |
Montana - University of Montana - Missoula Campus Department of Chemistry and Biochemistry, room 013, The University of Montana, Missoula |
| Manager or contact | Laurie Franklin |
| Users who may access |
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This listing was last modified on March 17, 2010.
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