This proposal requests funds for purchasing a Live Embryo/Live Tissue laser scanning confocal microscope. The Zeiss LSM710 live embryo/live tissue confocal instrument will be incorporated into the Cell Imaging Shared Resource (CISR) at Vanderbilt University, where it will allow users from across the Vanderbilt research community to perform long-term studies of 3-D embryogenesis and tissue morphogenesis. The Zeiss LSM710 confocal system detailed here is especially configured for long-term dynamic studies of living embryos and cultured tissues. Specifically, this system has increased sensitivity, allowing investigators to use reduced laser energy and exposure times. The increased sensitivity of this system allows visualization of living samples while minimizing damage from the excitation laser. A sophisticated live embryo/tissue incubator system will maintain exact physiological conditions with stable temperature, humidity, pH, and oxygen tension for long-term studies of embryogenesis and tissue morphogenesis. The specialized features of this confocal imaging system will therefore provide investigators with the ability to study the cellular and molecular events guiding 3-dimensional morphogenesis of multiple organs during development using new, state of the art approaches. The major user group consists of NIH-funded developmental biologists from multiple Vanderbilt Departments. Their specific research projects will use the Live Embryo Zeiss LSM710 confocal microscope to study embryonal and fetal development of multiple tissues and organs, including, foregut, choroid plexus, pancreas, lung, kidney, and heart. The flexibility of this system will allow studies using many different model systems from multiple species. As part of the Vanderbilt CISR, all major users will obtain expert training on the instrument and will have access to the instrument through the established CISR infrastructure. The Vanderbilt CISR manages a wide variety of microscopy resources for over 300 lab groups and receives outstanding institutional support from Vanderbilt University. Even with the extensive resources available in the Vanderbilt CISR, the Zeiss LSM710 fills a large void by allowing long-term studies of 3-dimensional morphogenesis. The inclusion of the Zeiss LSM710 in the Vanderbilt Cell Imaging Shared Resource will therefore provide NIH-funded investigators with a significantly more innovative approach to study important questions in developmental biology and tissue engineering.
