Research | Core Facilities | Patient Studies | Tech Transfer | Seminars | Intranet | Careers | Search | Contact Us | Ways To Give HOME
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Research | Core Facilities | Patient Studies | Tech Transfer | Seminars | Intranet | Careers | Search | Contact Us | Ways To Give HOME |
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Flow Cytometry and Cell Sorting Core Facility Introduction: The Flow Cytometry and Cell Sorting Core Facility supports many
OMRF investigators by providing training, analysis and multi-parameter, high speed cell
sorting. Mr. Jacob Bass and Dr. Diana Hamilton have completed the factory courses for these instruments and conduct all cell sorts for users. They also perform routine maintenance and data base archiving on the instruments and provide guidance on staining, sorting, special applications and analysis. In addition, they train postdoctoral fellows, graduate students and technicians in operation of the FACSCalibur, FACScan and LSR II. Instrumentation: Our DakoCytomation MLS3042 sorter purchased in 1998 is equipped with an Enterprise II 621 laser, a Coherent I70-4 laser and a 599-OEM Dye laser. It is routinely used for 2-way and 4-way sorts at speeds of up to 30,000 cells/second as well as single cell deposition sorts. In addition to green fluorescent protein, the most commonly used fluorochromes are FITC, PE, APC, Hoechst, PI, PE-TXRD, PE-CY5, and APC-CY5.5. A BD Biosciences FACSAria desktop cell sorter is also available for cell sorting. This instrument has no counterparts with respect to capabilities and ease of use. It utilizes 488 nm, 633 nm and 407 nm air-cooled lasers with advanced optics to detect up to 15 independent signals. The 488 nm laser can be used to detect common fluorochromes including GFP, FITC, PE, PI, and PE-Texas Red. The 633 nm laser is used to detect APC, APC-Cy7, APC-Cy5.5, and Cy5. These fluorochromes are most commonly used by our investigators who sort hematopoeitic progenitors or B cell subsets from tonsils. Finally, the 407 nm laser is appropriate for FRET experiments using cyan fluorescent protein. The LSRII analyzer is equipped with 488 nm and 633 nm lasers. This digital instrument is a good companion to the FACSAria and is capable of up to 12 parameters. A BD Biosciences FACScan and FACSCalibur cytometers are capable of three and four color fluorescence analyses, respectively. The Cyan flow cytometer offers cell analyses using a 488nm (blue), a 635nm (red), and a 405nm (violet) laser. Seven colors can be seen from the blue laser, two colors from the red laser and 2 colors from the violet laser. Summit software controls all aspects of the Cyan, such as cleaning cycles. A workstation connected to the Cyan also uses Summit software. To assist researchers with analysis, the Flow Lab has a Mac computer with Cellquest Pro and FlowJo and a PC with Diva and Summit. We also have three dongles for the Diva software that can be loaned. We also provide a transfer file on the sanserver and have CDs and DVDs available to make copies of data files. Even though we make backups of the computers, all users should frequently back-up their data.
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