Flow Cytometry/Cell Sorting - Services
Flow cytometry is a laser-based technology that permits rapid and precise multiparameter analysis of individual cells and particles from within a heterogeneous population as they flow in suspension past one or more laser beams. Using the different instruments in the shared resource, information on the physical and fluorescence properties of individual cells or organelles can be obtained at the rate of several thousand cells per second. Laser-based cytometry provides high intensity light sources for efficient and broad range illumination of fluorescent probes. The availability of multiple lasers on each of our instruments provides investigators with an opportunity to simultaneously measure several different characteristics of their cells. The instruments are linked to computer workstations with advanced software for rapid data analysis. Typically, investigators, their students/postdoctoral fellows and/or technicians work with technical experts in the shared resource to analyze samples and the resultant data, thereby gaining first-hand experience in data acquisition and analysis. The staff also works closely with investigators to design experiments and protocols for optimal utilization of the facility. A variety of different services currently available are listed below. Experimental protocols are available for each of these techniques to guide investigators. New protocols are routinely being established, tested and provided to investigators. Please contact shared resource facility staff for additional information.
Specific services include:
- DNA and RNA content
- Cell-cycle distribution and tumor ploidy determination
- Apoptosis assays (e.g., annexin V, TUNEL assay, caspase activity, PARP, etc.)
- Cell proliferation assays (e.g., Click-it Edu, CFSE, etc.)
- Cell function measurements (e.g., viability, oxidative metabolism, phagocytosis)
- Cell biochemistry (e.g., intracellular pH, calcium mobilization, mitochondrial membrane potential, microviscosity, glutathione content)
- Fluorescent protein expression analysis (GFP, YFP, RFP, mCherry, etc.)
- Chromosome number and identification
- Side population analysis for stem cell research
- Multiparameter cell sorting of up to four different populations simultaneously
- Multiparameter cell sorting for cloning and single-cell isolation into a variety of multi-well plates
A major focus of the Flow Cytometry/Cell Sorting Shared Resource is the development of training programs on instrumentation, applications, and data analysis. Training and education for independent use of the benchtop systems by graduate and undergraduate students, post-doctoral fellows, laboratory research staff and visiting researchers is provided regularly. These include individual user-based instruction, small-group training sessions, and tutorials. The latter format is more formal and has been quite successful in increasing the user base.