Flow Cytometry and Cell Separation Shared Resource: Project Summary Biology Shared Resource Group The Purdue Flow Cytometry and Cell Separation Shared Resource (FC-SR) provides advanced cell and particle analysis, and sorting capabilities to Purdue University Center for Cancer Research (PCCR) members. As an analytical tool, flow cytometry allows users to identify and quantitate cellular features, organelles, or structural components using optical measures. When combined with sorting capabilities, it is possible to identify and isolate single cells that have specific properties or express specific molecules and which can then be further examined and characterized. In cancer research, where the identification and characterization of unique, specific cells with cancer-like characteristics is critical, flow cytometry and cell sorting are extremely vital and useful to today's cancer researcher. Purdue's FC-SR provides a broad range of services that supported 24 Purdue Center for Cancer Research (PCCR) investigators' research projects from all four of PCCR's Research Programs in this current year. The FC-SR, located in the Bindley Bioscience Center (BBC) of Discovery Park, has grown considerably in both its technical expertise and instrument capabilities since its total re-organization in March 2009. A major emphasis of the facility is live cell sorting of mouse and human cells under BSL-2 conditions. In 2010 with support from an NIH S10 Shared Instrument Grant, the FC-SR obtained a high-throughput LEAP image scanning cytometer. This instrument adds an additional dimension to sorting of attached cells by laser ablation and serves as a bridge between flow cytometry and image analysis capabilities. The FC-SR also operates a FACS Aria III Sorter that serves as the primary workhorse for analysis and cell sorting. Currently, the facilty also operates a FC500 Cell Analyzer for routine cell analyses. The data analysis capabilities of the facility have advanced greatly with the addition of many FlowJo site licenses and the development of new advanced data analysis capabilities. Perhaps the most important resource provided by the FC-SR is the expertise that the staff shares. Many of FC-SR's clients have little, if any, experience conducting cytometry analysis or designing experiments using the BD FACS Aria III cell sorter. In the absence of the FC-SR and its experienced staff, these investigators would have tremendous difficulty in generating cytometry data necessary for competitive external funding.