Flow cytometry is a laser-based technology used to quantify specific properties of individual cells at a rapid rate. Prior to flow cytometric analysis, cells are prepared as a single-cell (monodispersed) suspension in a liquid buffer or medium. Depending upon the properties to be measured, the cells may be treated with one or more fluorescent reagent, whose binding to the cell is indicative of the cellular characteristic under investigation. During analysis, the cells are passed, single-file, past one or more lasers that are used to stimulate the fluorescence of reagents used. The intensity of fluorescence of each of the reagents used to treat the cells is detected and quantified by the flow cytometer for each cell that is detected. Usually, 10,000 cells are analyzed in less than a minute.
Cell properties that are typically analyzed by flow cytometry include, but certainly not limited to:
- Relative size
- Relative granularity
- DNA content
- RNA content
- Expression of specific cell surface molecules
- Expression of specific intracellular molecules
- Expression of fluorescent proteins
- Transient cell signaling events
In addition to the analytical flow cytometry described above, some of these instruments (cell sorters) are capable of physically separating selected subsets of cells from the original heterogeneous population for further growth or analysis.
The Flow Cytometry Lab of the Research Core Facility offers a very robust complement of flow cytometric services to investigators, including simultaneous analysis of up to 17 parameters and cell sorting of up to 4 cellular populations, again based on up to 17 parameters.
Matthew Woolard, PhD
Faculty Advisor, Flow Cytometry
Associate Professor, Microbiology & Immunology
Sumitra Miryala, PhD
Faculty Advisor, Cellular Metabolism
Assistant Professor, Cellular Biology & Anatomy