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Flow Cytometry Laboratory


Post-doctoral research associate preparing clonal cultures of Pfiesteria via cell sorting.

The Flow Cytometry Laboratory is responsible for cell quantification, sorting, and cloning. The lab uses a multi-laser research-grade COULTERŪ ALTRA™ flow cytometer, which is designed to pass individual cells or other small particles in single file through focused laser beams. The optical properties of each cell are measured as they pass through the laser, and a computer records these measurements. The flow cytometer allows the lab to rapidly count and identify different cell types in culture and field samples, based upon optical properties related to cell size, intracellular density, chlorophyll content, and fluorescence of applied molecular probes.

When certain cell types are identified, the flow cytometer can be used to physically separate specific individual cells away from the rest. Sorted cells can be collected in test tubes or on microscope slides for further culturing or examination. Cell sorting allows for very clean and pure cell cultures to be produced.

For example, toxic Pfiesteria zoospores are routinely purified and concentrated for use in refined toxicity assays (Figs. A & B)

A. Flow cytometric histogram generated from analysis of fish bioassay sample. Viable Pfiesteria spp. zoospores (quadrant A2) can be sorted out of fish bioassay water samples based upon optical properties related to particulate size and optical density.

B. Subsequent reanalysis of the sorted zoospores demonstrating the potential accuracy and precision of the sorting procedure. The sorted material is free of contaminants.

The flow cytometry lab is also equipped with a PHYTO-PAM™ Phytoplankton Analyzer, which employs pulses of colored light to measure the light absorption and fluorescence of phytoplankton cells in solution. These parameters can be used to quickly identify different types of phytoplankton present in a water sample. Photosynthetic efficiency can also be measured with this instrument, and directly related to the physiological health of the phytoplankton being analyzed. The lab also contains a Coulter Multisizer II Analyzer, which allows rapid counting and measurement of the population size distribution of cells in cultures and field samples. The graphically displayed results show cell number, volume, and surface area.

Flow Cytometry Laboratory at the Center for Applied Aquatic Ecology

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