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In‐vivo dynamic and hyperspectral microscopy

Microscopy techniques have advanced our understanding of healthy and diseased tissues for over 100 years. Over the past 20 years, techniques for in‐vivo microscopy have advanced, allowing examination of cells and tissues in situ, within the living body. While imaging in‐vivo presents many technical challenges, it also brings rewards. Imaging living tissues allows their function and physiology to be studied in real time, particularly their responses to stimulation or other interventions or therapies. These possibilities place new requirements on microscopy techniques to be able to capture dynamic events, in 3D within living tissues. Available contrast must also be considered. Substances suitable for specific labeling in‐vivo can be used, or imaging can visualize transfected cells expressing fluorescent proteins, some of which can report cellular function themselves. Additional contrast is available from intrinsic absorbers and fluorophores such as hemoglobin, NADH and FAD, whose dynamics can allow tissue metabolism to be directly visualized. Separating such sources of in‐vivo contrast, and moreover capturing multiple forms of contrast in parallel during dynamic cellular events can be achieved through hyperspectral microscopy. This talk will describe techniques optimized for in‐vivo microscopy, and will provide examples of the kinds of processes that can be visualized in the living brain and skin.