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 proliferation, and cytokine production.
Single and bulk cell sorting is performed based on expression of cell surface antigen(s) and/or expression of genetically engineered intercellular fluorescent proteins. The facility also provides instrumentation and expertise in single cell gene expression profiling.
This facility is currently equipped with three flow cytometers. The Becton-Dickenson (BD) FACSAria III is a 4 laser high-speed bench top sorter equipped with an automated cell deposition unit (ACDU) and housed in a biosafety cabinet for BSL2(+) sorting. The BD LSRFortessa is a 4 laser analyzer equipped with a high throughput sampling module, and a detector to analyze submicron-sized particles. The Cytek Aurora is a full spectrum 3 laser analyzer with a 96 well plate loader.
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 gene expression analysis, the facility has two instruments, the 10x Genomics Chromium Controller and the 10x Genomics Visium CytAssist. The 10x Genomics Chromium Controller instrument barcodes individual cell’s RNA for bulk single cell RNAseq and multiomic analysis including gene expression, cell surface proteins, antigen specificity, and chromatin accessibility. For spatial transcriptomics, the Visium CytAssist instrument can be used with fresh-frozen, fixed-frozen, and formalin-fixed paraffin-embedded tissue samples sectiononto standard glass slides. Transcriptomic probes can be transferred from slides containing tissue sections to obtain spatial data with gene expression.
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™ Pro located in a biosafety cabinet for sterile paramagnetic cell sorting and a Meso Scale Discovery Sector 2400 automated plate reader that detects multiple analytes by electrochemiluminescence.
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 2 print quality color laser printers.
The facility is located on the 2nd floor of the Robert C. Byrd Health Sciences Center North building, Rooms 2160 and 2184.
