Flow Cytometry and Immune Monitoring Core
Flow Cytometry and Immune Monitoring Core Facility
The University of Kentucky Flow Cytometry and Immune Monitoring Core Facility offers state-of-the-art flow cytometry cell analysis, high-speed sorting, and immune functional analysis to the research community at UK, and has been in continuous operation since 1983.
Our skilled staff provide a variety of flow cytometric approaches for multiparameter fluorescent analysis and sorting of eukaryotic and prokaryotic cell types and provide a variety of immune monitoring analyses to phenotype immune cell populations and functions. Cell sorting and analysis is also available for live human cells and infectious (BSL-2 level) samples. Training on analytical cytometers is available to investigators and their staff in order for laboratories to analyze their own samples at reduced cost.
Please do not hesitate to contact us if you have any questions about potential uses of flow cytometry for your research projects, how to appropriately design experiments for flow cytometry, prepare samples, or how to analyze your data.
Description of Core for inclusion in “facilities” section of grants
The Flow Cytometry and Immune Monitoring Core (FCIM) provides comprehensive, state-of-the-art analytical flow cytometry cell analysis and high-speed cell sorting services to support both human and animal studies. FCIM has been in continuous operation since 1983 and provides a variety of flow cytometric approaches for multiparameter fluorescent analysis and for the sorting of eukaryotic and prokaryotic cell types. Cell sorting and analysis is also available for live human cells and infectious (BSL-2 level) samples. Training on analytical cytometers is available to investigators and their staff so that individual laboratories can analyze their own samples at reduced cost.
Flow cytometric services include: 1) multiparameter cell phenotyping up to 23 colors on cell analyzers and 12 colors on cell sorters; 2) analysis of DNA content and cell cycle; 3) analysis of cellular apoptosis and necrosis; 4) quantification of cell cytotoxicity; 5) intracellular cytokine and protein analysis; 6) measurement of cell activation and signal transduction parameters; 7) high speed cell sorting of live human and animal cells under BSL-2 biocontainment conditions; and 8) single cell sorting into microtiter plates for cloning or single cell sequencing with next generation sequencing (NGS) technologies on campus.
Immune monitoring services include: peripheral blood cell isolation, plasma/serum isolation, cell banking, cytokine/growth factor analysis (ELISA, Elispot, Luminex) and leukocyte functional assays. In addition, FCIM also offers macrophage/DC generation, T cell expansion and other cellular product and immune assay design services.
Standardized data analysis is available to investigators for each set of samples, depending upon the application. Data can be reanalyzed by staff at the request of the investigator. The core also makes available several workstations and software options for users to analyze their own data at no additional charge. The workstations in the lab have either Mac-based CellQuest analytical software or PC-based Summit software as well as FlowJo software. A site license for FlowJo has been purchased and individual licenses are available to labs for a yearly fee.
Current instrumentation and equipment include the following systems and analyzers. A cell sorter system (iCyt-Sony) contains two separate sorter units housed in a BSL-2 level biosafety cabinet. Each sorter unit is equipped with 5 lasers and can sort or analyze up to 12 antibodies or fluorescent labels. A MACSQuant VYB (Miltenyi Biotec) with 3 lasers can measure up to 8 colors. A CytoFLEX LX cell analyzer (Beckman Coulter) with 4 lasers is capable of detecting up to 19 colors. A FACSymphony analyzer (Becton Dickinson) with 5 lasers can detect up to 23 colors. An LSR II analyzer is available which can detect 10 colors. A Magnetic Cell Separator (AutoMACS Pro) is available for bulk magnetic cell isolation.
Data Storage Policy
All flow cytometry data generated by FCIM is stored on a secure university server in a standard FCS format that is readable on any of the common analytical flow cytometry software programs. Researchers and their staff have ready access to any of their data generated over the previous 5 years via this server. The Flow Cytometry data on the network storage is backed up every night by IT Services to tape backups using IBM Tivoli Storage Manager software.