Illustrating Flow Cytometry in an Immunology Lab by Using a Hands-On Low-Tech Simulation

摘要

Flow cytometry is a foundational technique in immunology, microbiology, and other cell and molecular biology applications. The basic method involves passing suspended cells through a flow cytometer, which can detect and quantify the forward and side scattering of light passing through the cells. The cells can also be incubated with fluorescent probes or fluorophore-conjugated antibodies prior to passing them through the flow cytometer, allowing the presence and relative expression level of specific molecules to be analyzed. Cells can also be gated for the expression of specific molecules, and further analysis of other labeled molecules in the gated subpopulations of cells can then be performed. The data from a flow cytometry experiment is typically shown as dot/scatter plots or histograms. The cost of flow cytometry equipment has decreased substantially over the past few decades. However, it is still a major equipment purchase that is not available at many primarily undergraduate institutions, where flow cytometry-based experiments might typically be done in the context of a microbiology or immunology course rather than as a mainstay of a major research project. The cost of fluorescent-tagged antibodies and the availability of cells to label for a lab activity can also be barriers to doing flow cytometry experiments in an undergraduate lab course.