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 facility, information on the physical and fluorescence properties of individual cells or organelles can be obtained, as well as their subcellular localization. Laser-based cytometry provides a high intensity light source for efficient and broad range illumination of fluorescent probes (see Table 1 below). 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 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.
Specific services include:
- DNA and RNA content
- Cell-cycle distribution and tumor ploidy determination
- Apoptosis assays (e.g., annexin V, TUNEL assay, caspase activity, etc.)
- Cell proliferation assay
- 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)
- Chromosome number and identification
- Side population analysis for stem cell research
- Multiparameter cell sorting and cloning
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 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. In addition, this resource routinely gives classroom lectures and demonstrations for undergraduate and graduate students, as well as a formal course that students can take for credit.