Flow Cytometry and Single Cell Core

Overview of Services

The WVU Flow Cytometry & Single Cell Core Facility (FCSCCF) is a fee for service facility that 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 quantification, cytokine production, and cell sorting based on expression of cell surface antigen(s) and/or expression of genetically engineered intracellular fluorescent proteins.

The facility is equipped with four cytometers: a 4 laser BD FACSAria high speed bench top sorter equipped with an automated cell deposition unit (ACDU) for single cell sorting into plates, a 4 laser BD LSRFortessa with a high throughput sampling module and a detector to analyze submicron size particles, a 3 laser Cytek Aurora full spectrum analyzer with a 96 well plate loader, and a 1 laser Luminex Guava easyCyte HT System analyzer with a 96 well plate loader. For small particular detection (10 - 2000 nm), the facility has a NanoSight NS300 that detects, counts, sizes and determines, using fluorescently labeled antibodies, 1 - 4 proteins expressed by the particles

For single cell DNA/RNA analysis, the facility has a 10x Genomic Chromium Controller and a Fluidigm C1 Single Cell Auto Prep System. The 10x Genomics Chromium Controller barcodes individual cell’s RNA for bulk single cell RNAseq and multiomic analysis including gene expression, cell surface proteins, antigen specificity, and chromatin accessibility. The Fluidigm C1 Single Cell Auto Prep System, uses an integrated fluidic circuit chip to isolate 96 individual cells, extract the RNA/DNA/microRNA from each cell, amplify the RNA and make cDNA. The resulting products can be sequenced or used in PCR. 

Additional instrumentation in the lab includes:

 Miltenyi Biotec gentleMACS Otto Dissociator
For making single cell suspensions or homogenates from tissues

Miltenyi Biotec AutoMACS Pro
For sterile magnetic cell sorting

Zetasizer Nano Z
To measure the mobility and zeta potential of dispersed particles and molecules in solution

Meso Scale Discovery QuickPlex SQ120
Automated plate reader that detects multiple analytes by electrochemiluminescence

Personnel

Location and hours of operation

Links and Resources

1. http://flowcore.hsc.wvu.edu

Contacts