A replacement multiphoton microscope for in vivo imaging in rodent models of neur

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DESCRIPTION (provided by applicant): Multiphoton microscopy is a powerful technique for high resolution intravital imaging in deep tissue, with particularly exciting results obtained from brain imaging. We have successfully implemented and innovated in vivo brain imaging over the past 10 years, most notably in the study of Alzheimer's disease mouse models. These innovations, however, have extended to other brain imaging applications around the world where we have either directly or indirectly assisted in advancing the applications. Our application of multiphoton microscopy began with the first commercial multiphoton microscope, the Biorad 1024MP, which is still functional in the lab, but antiquated. There is no upgrade path for this instrument: it needs to be replaced. We are proposing to purchase a state-of-the-art multiphoton microscopy system to replace this instrument. The configuration of the microscope platform chosen is optimized for fast and deep in-vivo imaging. Using negative chirp optics to compensate for group velocity dispersion in the optical path, the proposed instrument allows optimized pulse-widths for deep tissue imaging with reduced average power. We have assembled a group of experienced, NIH-funded investigators with applications in a variety of neurological diseases that will take advantage of the capabilities of the instrument in an environment rich in experience and proven success. We have been using multiphoton microscopy to image structure and function in vivo on spatial scales as small as a single spine on an identified dendrite from a single neuron. We have developed approaches to image the pathology associated with Alzheimer's disease, neurons, glia, and the vasculature. We have also been using fluorescence lifetime imaging microscopy (FLIM) for sensitive detection of fluorescence resonance energy transfer (FRET), which can be used to reveal interactions between individual proteins. While FLIM has been largely relegated to in vitro or cell culture measurements, we aim to apply FLIM for quantitative imaging in populations of neurons and glia in vivo. The new microscope will be a vital resource for NIH funded imaging studies in mouse models of neurodegenerative diseases, stroke, and epilepsy. PUBLIC HEALTH RELEVANCE: We propose to replace an old multiphoton microscope with a newer more advanced model. We have been using multiphoton microscopy for 10 years to try to understand the pathophysiology in the brain of living mouse models of neurodegenerative disease, and a new microscope will ensure that we can continue our successful research programs.