Analytical Cytometry / Image Analysis Core Facility

Director: Debra Laskin, Ph.D.

Associate Director: Carol Gardner , Ph.D.
Associate Director: Vasanthi Sunil, Ph.D.
Laboratory Manager: Theresa Choi

Since 1985, the Flow Cytometry/Image Analysis Laboratory has been analyzing samples for academic and industrial researchers at reasonable costs. Flow Cytometry uses a laser-based instrument to analyze and sort cells or particles based on their light scattering properties and their pattern of fluorescence emission. Not only is the analysis fast, it yields information that is unattainable by other methods.

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 three 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. The goal of the Facility is to maximize the productivity of the users.

Confocal Microscopy

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.

Advanced software permits qualitative and quantitative analysis of images.