Multiphoton confocal microscope for in vivo biomedical imaging applications

Lo, David

Abstract

Confocal microscopy has enabled the detailed exploration of biological structures in three (and sometimes four) dimensions; aided by the expanding spectrum of fluorescent proteins and chemical fluorophores, interactions and relationships between proteins and cellular structures can be studied in greater detail. Multiphoton microscopy has become standard methodology in areas of biology where deep imaging of live tissue is required, and in chemistry and engineering where high energy and precise timing are required, but at present, no instrument at UC Riverside has multiphoton capability for in vivo imaging in the biomedical sciences. We propose to purchase a Nikon AIR MP+ high speed upright multiphoton system as the key instrument in a new In Vivo Imaging Core Facility in a Specific Pathogen-Free vivarium. Comparisons among competing systems showed the Nikon system to be the best match for our investigators' research needs, as well as providing the most functional capabilities such as spectral detection, within the budget allowed. Housing the multiphoton system in a procedure room adjacent to the vivarium will enable serial imaging in models of infection and inflammation, as well as in developmental models. To complement this instrument, our core group of microscopy users has assembled an extensive collection of fluorescent protein reporter transgenic and knockout mouse models for studying the interactions of proteins and cells in live tissues. These resources will be assembled as part of a dedicated facility to provide state-of-the art in vivo imaging instrumentation for the core users, as well as the biological research community on campus and at nearby research institutions. Strong institutional support and matching funds are provided by the UCR Office of Research and Economic Development and the School of Medicine. In addition, the majority of the core users are part of the Center for Glial-Neuronal Interactions, a research group supported by the School of Medicine that promotes interactive discussions and collaborations in Central Nervous System biology, with a key focus on microscopy imaging applications. The multiphoton microscope proposed for the In Vivo Imaging Core will be used in several in vivo models of neuronal development, in several types of studies on immunity and inflammation in the central nervous system, and in models of intestinal tissue biology. This core will be the center of an expanding in vivo imaging foundation that will build additional capabilities and develop innovative methods for following developmental and pathological processes in the brain and intestinal immune system, greatly expanding the capabilities available to researchers at UCR.