Flow Cytometry Facility

Services

1.  Multiparameter flow cytometry analysis and sorting services to basic research investigators, including operating the instruments or training investigators in assays such as:
a. Immunophenotyping/immunofluorescence analysis or sorting, up to 9 colors, using FITC, PE, and far red emitting dyes such as Red670, Red613, TriColor, PerCP, and ECD.  Other fluorochrom­es are also able to be used, including CY3, CY5, APC, and others.  PLEASE CHECK WITH FACILITY PERSONNEL CONCERNING WHICH FLUOROCHROMES AND COMBINATIONS WILL WORK.
b. Gene Expression and Transfection analysis or sorting, measured indirectly by using a reporter genes, such as Green Fluorescent Protein-type constructs (EGFP, etc.).
c. Cell cycle measurement.  DNA content, RNA content, and cell cycle profiling can be measured with PI, 7AAD,  or AO.  FITC conjugated anti-BrdU can provide additional information about cell cycle profiles when used along with PI.
d. Apoptosis.  Apoptosis/cell cycle assays are performed utilizing PI, 7AAD, FITC- or PE-UTP end labeling of fragmented DNA (TUNEL), Annexin-V, or AO in conjunction with EtBr.
e. Intercellular conjugation.  This assay has been run on cells stained with intracellular calcein, hydroxyethidine, or PKH26, as well as surface markers.  Total cellular protein can be measured with FITC or SR101.
f. Functional & kinetic assays.  Calcium flux and pH measurements can be performed utilizing ratiometric dyes, along with other available fluorochromes.
g. Cell Sorting.  Greater than 90% recovery of sorted populations, at consistent purity levels of >99%, at rates of 10,000 – 25,000 cells/second.

2.  Technical support in the form of protocol design assistance and data analysis required by the investigators.

3.  Training of investigators to allow them to independently perform data acquisition and analysis.

4.  Electronic storage, archiving, and retrieval of flow cytometric data through a local network and communication with the Institute network resource.

NO wet chemistry is performed in this facility.  Samples must be prepared by investigator and delivered to the facility, ready to be run on the flow cytometry equipment.