Flow cytometry is a laser-based technology that can rapidly analyze the biophysical, biochemical, and molecular properties of cells, most often through the use of fluorescent probes. Based on the fluorescent analysis of a mixed population of cells, a sub-population of cells that express a specific marker can be identified and physically sorted to a high purity. The laboratory has the ability to perform many advanced flow cytometric assays, including:

• Detection of cell-surface and intracellular antigens
• Cell cycle, cell division and DNA ploidy analysis
• Apoptosis assays
• Measurement of intracellular pH/calcium flux and uptake or efflux of fluorescent substrates
• Measurement of green fluorescent protein (GFP) and other fluorescent proteins
• High-throughput flow cytometric assays in 96-well plates

Laser	Wavelength (nm)	Commonly-used Fluorochromes
Coherent® Sapphire™ solid state	488 (blue)	FITC, PE, PE-Texas-Red, PerCP, PerCP-Cy5.5, PE-Cy7, PI
JDS Uniphase™ HeNe air cooled	633 (red)	APC, APC-Cy7, Cy5
Point Source Violet solid state	407 (violet)	Alexa Fluor- 405 and -430, Cascade Blue, Pacific Blue, DAPI, Hoechst

The flagship instrument of the laboratory is the Becton-Dickinson FACS Aria high-speed cell sorter. New-generation digital electronics allow high speed analysis and sorting (>20,000 cells per second). The FACS Aria optical system is configured with three excitation lasers and can detect 9 fluorescent parameters simultaneously. Additionally, the FACS Aria offers numerous sorting options which include four-way sorting, sorting to microtiter plates or glass slides, and customizable settings to optimize sort yield and purity. The FACS Aria laser excitation wavelengths and a partial list of fluorochromes that can be utilized on the instrument are listed in the table above.

For analytical purposes, the Becton-Dickinson LSR II flow cytometer is available. The LSR II has digital electronics for increased speed, a three-laser optical bench (see table above), and can detect eight fluorescent parameters (plus two physical parameters and a time parameter).

Guava Technologies PCA-96 is a table-top analytical flow cytometer with a user-friendly interface for single-color analysis of small samples. This instrument is pre-programmed for several common assays, which can be carried out in 96-well plates, allowing for high-throughput flow cytometric analysis.

The laboratory also has a dedicated workstation for data analysis and the preparation of graphics. FACS Diva, WinMDI, and ModFit LT software packages are available for such work.

For further information about the Flow Cytometry Core’s capabilities, technical considerations in experimental design, fee schedule, or to make an appointment to use the instrumentation, please contact.