Services and Fees
Flow Cytometry and Cell Sorting
Modern methods of flow cytometric cell analysis, with high-speed data acquisition and mass storage of data allow simultaneous measurement of up to 12 parameters on a single cell. They provide indispensable tools for the study of cell proliferation and differentiation pathways, gene regulation, tumor cell lineage, apoptosis and residual disease. When coupled with cell sorting, the information derived from multiparameter analysis is amplified significantly by the ability to further characterize the sorted cells with modern techniques of molecular biology, such as PCR and FISH. The following established assays have been tested and are available through the Core Facility:
Flow Cytometry
- Cell cycle and DNA ploidy analysis: EB, DAPI, BUdR, IUdR, Ki67, PCNA, AO, nucleolar antigens, cyclins
- Apoptosis: TUNEL, subG1 (PI), phosphatidyl serine/annexin V, caspase activation, mitochondrial membrane potential (CMXRos), Bcl-2, Bax, Bcl-X, BAG-1, Fas, CD30, CD40
- Ca++ mobilization
- Differentiation: lineage/differentiation specific cell-surface antigens
- Cell division history: PKH26 allows determination of the number of actual cell divisions in vitro and sorting of quiescent and proliferating cells for molecular analysis
- Transgene detection: GFP, beta-gal, NGF-R, MDR1
- Receptors: fluorescence-tagged cytokines, IL-2R
- Oncogenes: p53, c-myc, p21, ras
- Drug resistance: MDR1, LRP, BCRP, drug uptake and efflux
Cell Sorting
- Chromosome sorting
- Hematopoietic progenitor and stem cell sorting (e.g., CD34+/38-, CD34+/38+, “side population” [SP] cells, which eliminate Hoechst 33342
- GFP-expressing transfected cells
- All other assays listed above can be the basis for FACS sorting
Laser Scanning Cytometry
This is a slide-based system with fluorescence detection that can analyze up to three fluorochromes from cells deposited on slides. FACS sorted or rare tumor cells can be further analyzed using this technology. The LSC is the only system capable of determining protein quantitation in single cells.
Laser Scanning Confocal Microscopy
This process allows the determination of localization and co-localization of cellular and sub-cellular components. The Core Facility can offer researchers tools and techniques for image acquisition, 3D-reconstruction and time-series observation as well as a variety of image processing and analysis functions.
Charges
- $50/hour for Flow Cytometry and Cell Sorting
- $50/hour for Confocal Microscopy