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Analytical Cytometry / Image Analysis

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Services

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 the 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 new laser-scanning confocal microscopes will provide three-dimensional images on various structural, biochemical, and functional aspects of tissues and cell 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 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 and tested.

Specific services include:

  • FACS Based Analysis of Cell Populations
  • Cell Sorting and Cloning
  • Cell-Cycle Distribution
  • Analysis of Green Fluorescence Protein Expression
  • Apoptosis Assays
  • DNA and RNA Content Analysis
  • Tumor Ploidy Determination
  • Immunofluorescence Microscopy
  • Cell Function Measurements (i.e., Oxidative Metabolism, Phagocytosis)
  • Cellular Biochemistry (i.e., Intracellular Ph, Calcium Mobilization, Membrane Potential, Microviscosity, Glutathione content)
  • Chromosome Number and Identification
  • Fluorescence Confocal Microscopy
  • Fluorescence in situ Hybridization (FISH)
  • Fluorescence Image Analysis of Individual Cells and Tissue Samples
  • Cell-Cell Communication and Fluorescence Redistribution After Photolabeling (FRAP)

A major focus of the shared resource is the development of training programs on instrumentation, applications, and data analysis. 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.