This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Recent developments in optical imaging have revolutionized the way in which we can examine cells and tissue in vivo. The number of modern optical imaging technique is continuously expanding, each offering a unique contrast mechanism and sensitivity to important bio-compounds in the system under study. For instance, tissue morphology can be non-invasively examined with optical coherence tomography (OCT). Over the past decade, the development of nonlinear methods such as two photon excited fluorescence (TPEF), second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) microscopy has given the biomedical researcher convenient tools for selectively visualizing endogenous structures without the need of labeling. The multi-dimensional approach to microscopy integrates various imaging techniques and contrast mechanisms to better assess the biological specimen. Combinations of the TPEF, SHG, CARS and OCT techniques have previously been realized into optical microscopes, however, a full integration of these imaging modalities has not been accomplished. In this proposal we push the envelope of multi-dimensional imaging in the following way: a)Combining the TPEF, SHG, CARS and OCT into a single imaging platform. b)Optimizing the image contrast and penetration depth by controlling the spectral phase. The proposed multi-dimensional imaging platform uniquely permits the three-dimensional mapping of key tissue and cell components such as flavin metabolites, elastin networks, collagen filaments and lipid pools along with the detailed tissue morphology. Such an optical assessment of the biological specimen would provide one of the most detailed microscopic views of the sample currently possible.
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