The scientific community of the University of Pennsylvania School of Medicine has many collaborative and independent areas of investigation which will benefit from the new fluorescence activated cell sorter (FACStar Plus). While the range of user interests is evident in the diversity of the proposed projects, each bears important implications for understanding and treating disease as well as providing insight into basic processes of cell activation and differentiation. The diversity of the proposals is indicative of the need for a highly versatile, state of the art, flow cytometry machine. The FACStar Plus will provide the greatest flexibility and state-of-the art capabilities currently available for both cell sorting and analytical functions. The proposed configuration will contain two lasers and seven detectors allowing forward and orthogonal light scatter measurements along with five fluorescence detectors. The combination argon-ion and helium-neon (HeNe) laser system will provide us with the most cost- effective flexibility in analytical flow cytometry and cell sorting currently available. A Coherent 5 watt argon-ion laser has multiline visible output lines from 457.9 to 514.5 nm and multiline UV output lines from 333.6 to 363.8 nm at 400 mW. The blue 488 nm output line will allow the use of FITC, PE, propidium iodide and a choice of several available tandem dye conjugates. The 363 nm UV output line will allow the use of the calcium indicator Indo-1 or the DNA binding Hoechst 33342 and DAPI. The air-cooled helium-neon laser will provide 35mW output at 633 nm. This will allow the use of the phycobiliprotein allophycocyanin (APC) and potentially the use of a new family of fluorochromes such as cyanin-5. When combined with powerful data management provided by a Hewlett-Packard 340 workstation and the LYSYS II software, this equipment will provide the versatility, modularity and open-ended design necessary for advanced flow cytometric applications.
