The Flow Cytometry and Cell Sorting Facility at the Abramson Cancer Center (ACC) is recognized as a worldwide leader in cytometry and cytomics and is one of the largest and most comprehensive academic flow cytometry Shared Resources in North America. A full range of cytomic services is offered including analytical cytometry of up to 30 parameters, detection and analysis of extracellular vesicles, basic and ultra-rare cell sorting, single cell multiomic analysis, computational cytomic data analysis, permanent data archive, expert consultation, and a comprehensive education program. Jonni Moore, PhD, Professor of Pathology and Laboratory Medicine with a doctorate in immunology, has directed the Facility since 1990.
The services provided by the Flow Cytometry and Cell Sorting Facility can be divided into several areas:
- Direct technical services
- Technical consultation and experimental design
- Education and training
Several new services and instruments were added in the last several years (indicated by *).
Direct Technical Services
Typical services include, but are not limited to:
- Investigator Performed Services (with consultation, training and support of facility staff as needed):
- *High parameter immunophenotypic applications (surface and intracellular, up to 30 parameters)
- Functional probes and assays (membrane integrity and potential, cell viability and proliferation, signal transduction, enzyme substrate/ligand binding studies, reactive oxygen species, calcium flux, fluorescence resonance energy transfer, cell tracking, DNA content and cell cycle, apoptosis, etc.)
- Imaging cytometry (Amnis Imagestream)
- Cell sorting on FACS Aria, FACS Jazz, and *FACS Melody for up to 6-way sorting and 16 parameters
- *Multiomic RNA-seq/Ab-seq applications using BD Rhapsody on site in proximity to sorting
- Facility Performed Technical Services
- Up to 6-way sorting and single-cell index sorting, including infectious sorting in a BSL2 setting
- Sixteen parameter sorting (with blue, red, green and violet excitation) with variable speed and pressure customization for optimized recovery and viability
- *Novel facility developed applications for extracellular vesicle detection with a specially modified FACS Canto II and custom computational methods
- Computational modeling and advanced data analysis; permanent cloud-based data archiving; site-licensing of analysis software
- New assay or application development based on investigator needs including *translational applications support with GLP-like validation
- Special sorting with FACS Focus for enrichment of ultra-rare cells
Technical Consultation and Experimental Design
Consultation offers expert cytometry support for the design of simple to highly complex experiments by researchers, increasing the success rate of ACC investigator studies. *A dedicated PhD-level Technical Director for Research and Development, Dr. Derek Jones, was added in 2017 to further enhance the consultation service.
Education and Training
A full curriculum of educational services, ranging from formal classes to workshops to lab practicums and individual tutorials, is available. This enables ACC members, their trainees and staff, regardless of their level of experience, access to virtually all applications of flow cytometry/cell sorting. More than 4,000 new users have been trained in the fundamentals of flow cytometry since 1996. Training is easily accessed and provided frequently so that new investigators have access to the technology very quickly.
Technologies and Major Equipment
As of September 2019, the lab operates or has access to 32 instruments:
- 8 cell sorters (2 BD Influx, 4 BD FACS Aria, 1 BD FACS Jazz, *1 BD FACS Melody; 6-16 colors)
- 12 standard analyzers (5 BD LSRII, 1 BD FACS Canto, 1 BD Accuri, 1 BD FACS Lyric, *(2 new) 4 BD LSR Fortessa; 4-18 colors)
- *4 High Parameter Analyzers (3 BD FACSymphony A5, 1 BD FACSymphony A3-Lite; 23-28 colors)
- 3 specially modified FACS Cantos (10-11 color)
- *1 unique extracellular vesicle analyzer (FACS CantoMP, high intensity laser, optics modified; 10 color)
- 1 Coulter Vi-cell
- 1 Amnis ImageStream
- *1 single cell gene expression analysis system (BD Rhapsody)
- 1 CyTOF mass cytometer (collaboration with Philadelphia VA Hospital across the street)
In addition, biocomputational services for the modeling and analysis of data are offered by the embedded biocomputational group. Using facility-designed systems, ALL data collected on all instruments are permanently archived offsite and are easily retrievable when needed.
For more information, please visit the Facility's website: https://pathbio.med.upenn.edu/pbr/portal/flowcyto