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| Hudson-Webber Cancer Research Center rooms 812 & 823, 4100 John R, 966-2687 |
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Prentis Cancer Center room 426, 110 E. Warren, 833-0715 x2365 |
The all-digital FACS Vantage SE/DiVa sorts cells at up to 16,000 cells per second into test tubes, multiwell culture plates, or microscope slides. It can excite a variety of fluorochromes with 3 lasers: 488 nm, multiline UV (351-364 nm), and rhodamine 6G dye laser (tunable to 580-620 nm). The FACS DiVa is our only cytometer capable of measuring UV-excited fluorochromes (indo-1, Cascade Blue, AMCA, DAPI, Hoechst 33342, Calcofluor White, etc.), and "yellow"-excited fluorochromes (Texas Red, sulforhodamine 101, etc.). The 488 nm laser excites the more common fluorochromes used in flow cytometry: fluorescein isothiocyanate (FITC), green fluorescent protein (GFP), R-phycoerythrin (PE), propidium iodide (PI), and PE-Cy5 tandems (Red670, CyChrome, TriColor, PC5). Up to 4 fluorochromes can be excited simultaneously by the 488 nm laser; usually these are FITC, PE, PE-Cy5, and PE-Cy7. The UV laser can detect two fluorochromes simultaneously; one or both may be a Quantum Dot (Qdot). The dye laser can excite allophycocyanin (APC) and an APC-Cy7 tandem simultaneously. It is also possible to adjust the lasers for simultaneous UV and 458 nm output, allowing excitation of Hoechst 33258 and chromomycin A3 for flow karyotyping and sorting of chromosomes. The FACS DiVa is manufactured by BD. top
The FACSCalibur has 2 lasers emitting at 488 nm and ~635 nm. Up to 4 fluorochromes can be measured simultaneously; usually these are FITC, PE, PE-Cy5 (or PerCP), and APC. The red diode laser has also been used to excite TO-PRO-3, a nucleic acid dye typically used to label DNA. The FACSCalibur is manufactured by BD. top
The FACScan only has a 488 nm laser, but it can excite the most common fluorochromes used in flow cytometry. Up to 3 fluorochromes can be measured simultaneously; usually these are FITC, PE, and PE-Cy5 (or PerCP). DNA content measurement for cell cycle analysis is usually done with PI-stained samples. Other common assays run on the FACScan are Annexin V-FITC/PI for apoptosis and GFP for gene expression. Oxidative burst in PMN cells has been measured with 2,7-dichlorofluorescein diacetate (DCF). The FACScan is manufactured by BD. top
The LSC has both 488 nm and 633 nm lasers. Although 4 fluorescence detectors are dedicated to each laser (8 detectors total), it is not practical to use them all simultaneously. The LSC measures cells on a microscope slide, and can image selected cells using bright field or epifluorescence microscopy. Although the LSC can detect cells by laser scatter, it is preferred to use a fluorescent parameter for cell detection, such as PI-staining for DNA content. The LSC is manufactured by CompuCyte. top