Frederick CCR Flow Cytometry Core
Cancer and Inflammation Program (CIP)

Comprehensive support for studies of cell immunophenotyping, cell sorting, cell cycle, apoptosis and cell proliferation

Mission: Flow cytomety (FCM) is a unique experimental technology, which provides rapid, quantitative, multi-parametric, single cell analysis and separation. The mission of the flow cytometry core encompasses basic research support provided to members of the Frederick-CCR community. The laboratory staff also participates in the development of flow cytometry applications and resources for use in basic research. Resource development in the lab includes advances in instrumentation, methodology and computer/software. The core staff also trains investigators to operate the analyzer instruments. This laboratory is contract-operated with oversight by a user based steering committee.

Access: Dedicated to CCR and Program

Staff:
CCR Core Supervisor - Kathleen Noer, BS noerk@mail.nih.gov 301-846-5811
Support Staff - Roberta Matthai, BA matthair@mail.nih.gov 301-846-5811
                     - Guity Mohammadi, BS mohammadig@mail.nih.gov 301-846-5811

Expertise:
The CCR Flow Cytometry Core provides multi-color flow cytometry analysis of live or fixed single-cell suspensions derived from various tissues and cell cultures. The group can offer up to 12 parameter cell surface markers coupled with calcium-flux or cell cycle analysis. Standard and high-speed cell sorting of single-cell suspensions of up to 12 colors is available in either bulk sorts or by single cell deposition in 96-well plates. Cell cycle analysis software is available for cell ploidy, cell cycle, and apoptotic analysis of cell populations.

Established Technologies:

• Immunophenotyping of mouse and human cells using up to 17 parameters
• Apoptosis assays using annexin V, tunnel, caspase activation and mitochondrial membrane potential changes
• Cell cycle analysis and sorting
• Cell sorting using up to 12 colors
• Phenotyping of cells in proliferation assay
• Sorting and analysis of cells expressing multiple fluorescent proteins

Instrumentation:

• BD FACSAria II SORP cell sorter with five lasers (excitation lines at 488nm, 647nm, 405nm, 561nm and 355nm), 17 fluorescent detectors, high speed four-way bulk sorting and single cell deposition sorting
• BD FACSAria II cell sorter with three lasers (excitation lines at 488nm, 633nm and 405nm), 9 fluorescent detectors and high speed four-way cell sorting
• Beckman Coulter Cytomation MoFlo cell sorter with three lasers (excitation lines at 488nm, 405nm, and a Krypton tunable to 647nm and MLUV), 8 fluorescent detectors, high speed four-way bulk sorting and single cell deposition sorting
• BD LSRII SORP analyzer with four lasers (excitation lines at 488nm, 405nm, 561nm and 647nm) and 19 fluorescent detectors
• BD LSRII analyzer with three lasers (excitation lines at 488nm, 405nm and 633nm) and 9 fluorescent detectors
• BD Canto II analyzer with three lasers (excitation lines at 488nm, 405nm, and 647nm) and 8 fluorescent detectors
• BD LSRII Fortessa analyzer with two lasers (excitation 488nm and 647nm) and 6 fluorescent detectors.
• BD FACSCalibur analyzer with two lasers (excitation lines at 488 nm and 635nm) and four fluorescent detectors
• Two BD FACScan analyzers with one laser excitation line at 488nm and three fluorescent detectors

Collaborations:
Part of the mission of the group is to give advice and experimental design suggestions to the investigators who use this facility. This advice includes trouble-shooting when results do not meet the expectation of the investigator. Staining protocols are kept on file and are provided to the investigator.