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 of up to 18 colors or sorting up to 9 colors, using commonly available fluorochromes for immunofluorescence.  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. Maintenance and many repairs of all flow cytometer instrumentation, allowing for maximum uptime, in addition to tremendous saving in service contract costs.

5. 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.