A FACS Vantage TM cell sorter is being requested in this shared instrumentation grant. It will be operated in a core facility and serve a research foundation of some 100 scientists. This institution is currently supported by 35 NIH grants and a number of those projects are highly dependent on cell sorting. Additional scientists and projects utilize analytical flow cytometry and some of those would benefit from UV laser excitation provided by the Vantage TM. The needs being addressed by this application include routine four color flow cytometry and sorting, single cell sorting for cloning, intracellular [Ca++] measurements, and cell cycle determinations, as well as sorting of viable cells based on this parameter. We plan to configure this instrument with a water cooled, argon UV laser (Coherent Enterprise II) as well as an air cooled helium-neon laser (Spectra Physics). There will be detectors for five fluorescence channels in addition to ones for forward and side scatter. To facilitate DNA and other analyses, the instrument will have a pulse processor unit. Single or prescribed numbers of cells can be sorted with an automated deposition system while the viability and physiologic condition of cells will be maintained with a refrigerator-heater system. Because of the heavy sorting applications required of this instrument, we plan to add a TurboSort option to effectively double the sort rate. Data analysis will be facilitated by links to other computers and an analytical flow cytometer (FACScan) equipped with the same software. A connection to our Digital VAX computer will permit tong term data archival. There is currently no cell sorter in the Oklahoma Medical Research Foundation.
