This shared equipment proposal seeks to acquire a state-of-the-art fully automated perifusion system at VA Puget Sound Health Care System (VAPSHCS. The proposed instrument (a Biorep Technologies PERI4- FA-115 perifusion system) comprises an integrated perifusion system which simultaneously delivers solutions to multiple cell/tissue samples under highly controlled temperature, time and flow-rate conditions. The system also includes an integrated, robotic sample collection module, which allows collection of up to 36 samples simultaneously, and up to 1,920 samples per experiment. Finally, the system is fully automated, providing significant increases in efficiency and time-savings over conventional approaches for studies of this kind. This instrument will be housed, managed and maintained within the Cell Function and Analysis Sub-Core which is located at the Seattle Division of VAPSHCS. The proposed equipment purchase expands VAPSHCS?s research infrastructure, meeting an unmet need for the research program (described below). Specifically, there is no instrument on station at present that is capable of this functionality. Further, there is only one similar instrument within the Seattle area. This instrument is heavily used and located several miles away, precluding its use for time-sensitive studies for which this instrument is designed. Thus, investigators at VAPSHCS currently have no capability to perform the studies supported by the proposed perifusion system. The Biorep perifusion system enables real time, in vitro analysis of cell or tissue samples in response to a range of stimuli. This system is amenable to almost any cell or tissue type and is ideally suited for studies looking at dynamic changes in the production of secreted analytes or changes in cell viability. Importantly, it is widely applicable to our diverse community of researchers which includes those working in diabetes, cancer, obesity, liver disease, traumatic brain injury and neurodegenerative diseases. Twelve investigators on station working across these areas have an immediate need for the proposed instrument. Nine of these VA-funded (using ~75% of instrument time). These include 6 Major users (whose usage will total 28 h/week). The 3 remaining VA-funded investigators together with 3 additional users (with funding by agencies such as NIH), comprise the Minor user group (6 total; usage of 16 h/week. We present a comprehensive plan for the sharing, administration, scheduling (including use outside of regular business hours) and care of the instruments. A summary of some of the different uses of this system is given here (and detailed in the Research Plan): Assessment of dynamic insulin release from pancreatic islets in response to a variety of physiological stimuli, using samples from normal or diabetic humans or animal models Determination of inflammatory molecule production from vascular cells, leukocytes, liver, muscle and adipose tissue specimens under normal and disease conditions Evaluation of neuropeptide release from brain slices Measurement of cell viability in response to stressors in all of the above cell types In summary, this new instrument package will provide a state-of-the-art scientific platform to support many investigators based at VAPSHCS in Seattle. The instrument will be located and managed by the established Cell Function and Analysis Sub-Core, which already includes well-developed administrative and technical infrastructure for the management, scheduling and support of the Core?s services.
Real-time analysis of parameters related to cellular function and viability provides fundamental information about normal tissue and what changes occur in disease. Accurate and reliable data from these analyses is only possible when cell and tissue specimens are studied under optimal conditions. The ability to achieve rapid reproducible changes in the cellular environment is key to the ability to study these parameters. This request for a Biorep Perifusion System, which is fully automated, will significantly enhance the research capabilities of investigators at VA Puget Sound Health Care System who are working to improve the health of our Veterans. The group of major (6) and minor (6) users (comprising 9 VA-funded investigators) who will use this instrument are studying diabetes, cancer, obesity, liver disease, traumatic brain injury and neurodegenerative diseases. Each of these areas of research is an important focus for Veteran?s health, and includes many service- connected diseases.
