We are requesting a state of the art laser capture microscope (LCM) as the seminal instrument for an institutionally supported LCM core facility designed to service a broad spectrum of biological disciplines within the University of Connecticut. Our University is experiencing a major infusion of capital and operational funds from the State as part of the concept called Bioscience Connecticut. An immediate consequence of this investment was the decision by the Jackson Laboratories to establish a major research effort in the genomics of human cancer. This UCONN/JAX affiliation extends to their mouse genetic research programs located in Maine. To meet the increasing demands of this well-funded and broad user group, the instrument, Arcturus model XTTM, will be capable of immunofluorescence-based imaging that builds on the foundation for PCR analysis of laser captured samples developed by the core director, Dr. Joel Pachter. This particular model is unique among all LCM instruments currently available in having both infrared and ultraviolet laser capacities, providing users the greatest flexibility in cell acquisition. The technology willbe extended to capture cells within tissue or cell culture expressing GFP and a variety of fluorescence-based enzyme stains from which an extensive RNAseq analysis can be performed using an Ion Torrent platform. Capabilities for target capture based on multiplexing fluorescence signal and computer defined target selection will allow for flexible growth as new tissue sources are analyzed. The core will train users in the operation of the instrument and introduce them to a workflow that includes extensive RNA expression and bioinformatics characterization. The financial and compliance aspects of this fee for service core will be handled by existing reagent and common instrument core (Molecular Store) directed by the co-director of the LCM core (D. Rowe) using a server controlled software program that is easily adapted to activities of the LCM core. An experienced advisory committee has been established to ensure optimal utilization and attention to emerging applications of the instrument. In the process of implementing our LCM core concept, we will become a Life Technology Center of Excellence for our concept of linking LCM and RNAseq. This designation identifies us as a model for programs outside our University, and enables us to be involved in the development and implementation of new applications of the LCM platform. We have the fundamentals of institutional support, technical expertise, window to new applications, and the operational and financial experience in place to provide a heavily utilized and rapidly evolving core facility that will support even greater succes of our research faculty for funding and program development.
