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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 two flow cytometers.  The Becton-Dickenson (BD) FACSAria III is a 4 laser high-speed bench top sorter. The BD LSRFortessa  is a 4 laser analyzer equipped with a high throughput sampling module, and a detector to analyze submicron-sized particles. For submicron particle analysis, the facility has a Malvern NanoSight NS300 and a Zetasizer Nano Z. The NanoSight NS300 detects, counts, and measure the size of particles. It is equipped with 4 lasers for detection of fluorescently labeled particles. The Zetasizer Nano Z measures the zeta potential (the charge on particles in a solution) via electrophoretic mobility of particles in a solution using laser Doppler microelectrophoresis.

For single cell analysis, the facility has 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, and BD FACSDiva software for flow cytometry data acquisition and analysis. It also has a 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.

 

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