Analytical cytometry is a laser-based technology that permits rapid and precise multiparameter analysis of individual cells and particles from within a heterogeneous population or tissue section. Using the instruments in this facility, information on the physical and fluorescence properties of individual cells or organelles can be obtained, as well as their tissue and subcellular localization.
The Analytical Cytometry/Image Analysis Shared Resource provides investigators with rapid and precise multiparameter analysis of cells, particles, and tissue samples. Laser based cytometry provides a high intensity light source for efficient and broad range illumination of fluorescent probes.
The instruments are linked to computer work stations for rapid data analysis. Typically, investigators, their students, 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 methods are routinely being established and tested.
- DNA and RNA content
- Cell-cycle distribution and Tumor-ploidy determination
- Apoptosis assays (Annexin V, TUNEL assay, and Caspase activity, etc.)
- Cell proliferation assay
- Cell function measurements (i.e., viability, oxidative metabolism, and phagocytosis)
- Cell biochemistry (i.e., intracellular pH, calcium mobilization, mitochondria membrane potential, microviscosity, and glutathione content)
- Analysis of fluorescent protein (GFP, YFP, and RFP) expression
- Side population analysis for stem cell research
- Fluorescence imaging of individual cells and tissue samples
- Cell sorting and cloning
- IVIS Small Animal Imaging
The Facility is also equipped with a laser-based scanning confocal microscope (Leica TCS SP5). This instrument can be used to:
- Conduct advanced imaging of tissue sections, cell monolayers and single cells.
- Quantitate Ca++, pH, and membrane potential transients of viable cells.
- Conduct fluorescent in situ hybridization (FISH) analysis in tissue sections or single cells.
- Collect 3-D information from complex samples/reconstruct 3-D images for extended analysis.
- Image subcellular processes as they occur.