The Flow Cytometry/Cell Sorting & Confocal Microscopy Core Facility has been analyzing samples for academic and industrial researchers at reasonable costs since 1985. The goal of the Facility is to give investigators access to state-of-the-art instrumentation, technical expertise and experimental protocols for analysis of cells, particles, and tissues by flow cytometry / cell sorting and confocal microscopy.
Flow cytometry is used extensively for immunophenotyping of circulating white blood cells. First, blood leukocytes are resolved on the basis of size and granularity. The subpopulations of interest are then gated and analyzed for the presence of specific cell surface antigens using antibodies conjugated to fluorescent probes. This technology can be applied to any cell type to analyze extracellular or intracellular proteins. Cells of interest can also be separated (sorted) from the mixed populations and cultured for further analysis.
Various biochemical and functional properties of cells can be analyzed including hydrogen peroxide production, calcium mobilization, membrane potential and intracellular pH. Cellular processes associated with multi-drug resistance of tumors can be characterized using this technology.
Many pharmacologically active agents alter the growth properties of cells. Detailed analysis of the effects of these agents on cell cycle can be conducted in both fixed and viable cells. Mechanisms of apoptosis can be investigated as well as cell cycle dependent expression of specific proteins. Cellular DNA and RNA content can also be simultaneously measured.
Cytometric analysis is not limited to the study of mammalian cells. This technology can also be applied to lower eukaryotes (i.e. yeast) as well as bacteria. Cell cycle kinetics and various aspects of metabolism can be studied. Cells of interest can be sorted and clonally propagated.
The Cytometry Facility currently has two flow cytometers. These instruments are user-friendly and standard protocols are available for various applications. Following a brief tutorial, investigators/students analyze their samples, independently.
Cytometry is a process in which physical and/or chemical characteristics of single cells, or by extension, of other biological or nonbiological particles in roughly the same size range, are measured. In flow cytometry, the measurements are made as the cells or particles pass through the measuring apparatus, a flow cytometer, in a fluid stream. A cell sorter, or flow sorter, is a flow cytometer that uses electrical and/or mechanical means to divert and collect cells (or other small particles) with measured characteristics that fall within a user-selected range of values.