The WVU Flow Cytometry & Single Cell Core Facility (FCSCCF) provides instrumentation and scientific support for single cell analysis and sorting. The facility routinely performs analysis of both eukaryotic and prokaryotic cells for expression of intracellular and extracellular proteins, cell cycle, cell quantitation, cytokine production, and cell sorting based on expression of cell surface antigen(s) and/or expression of genetically engineered intercellular fluorescent proteins.
This facility is currently equipped with three cytometers. The Becton-Dickenson (BD) FACSAria is a high-speed bench top sorter. One of the analytical flow cytometers in the lab is the BD LSRFortessa which has 4 lasers, a high throughput sampling module, and a detector to analyze submicron-sized particles. The other analyzer in the facility is a BD dual laser FACSCalibur equipped with a sample autoloader and has the capability for 4 color analysis.
The facility has recently acquired a Fluidigm C1 Single Cell Auto Prep System. This instrument uses an integrated fluidic circuit (IFC) chip to isolate 96 individual cells. While in the IFC, RNA/DNA/microRNA is extracted from the single cells followed by amplification and cDNA generation (if needed). The resulting products can be sequenced or used in PCR. Other instrumentation in the facility include a Miltenyi Biotec gentleMACS™ Otto Dissociator for disassociation of tissues into single cells or homogenization for molecular biology applications, a Miltenyi Biotec AutoMACS™ located in a tissue culture hood for sterile magnetic cell sorting and a Meso Scale Discovery Sector 2400 automated plate reader that detects multiple analytes by electrochemilumiscence.
The facility has auxiliary laboratory space with appropriate equipment to train personnel and support the needs of researchers. The FCSCCF has several software packages including FCS Express, CellQuest Pro and BD FACSDiva software for flow cytometry data acquisition and analysis. It also has two separate analysis work stations with 3 print quality color laser printers.
The facility is located on the 2nd floor of the Robert C. Byrd Health Sciences Center North building, Room 2160.