The Flow Cytometry and Cell Sorting Core Facility supports many OMRF investigators by providing training, analysis and multi-parameter, high speed cell sorting. The facility is housed in Room E402.
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.
Contact Jacob Bass or Diana Hamilton at (405) 271-7299 or (405) 271-7261 or by email at bassj@omrf.org or hamiltond@omrf.org.
User name is flowlab and the password is cytometry. The FacsAria and MoFlo are cell sorters and can be scheduled two weeks in advance. The FacScan, FacsCalibur and LSRII are cell analyzers and can be scheduled one month in advance. Users can edit and delete their times for the analyzers but not for the sorters. Cancellations for the Aria and MoFlo should be made 24 hours in advance; otherwise, there is a one-hour cancellation charge.
Trained users who feel confident to use our analyzers after hours (after 5 p.m. on the weekdays and the whole weekend) may do so. Any user that signs up before 9:30 a.m. on the LSRII should turn the machine on, since it requires a longer warm-up period.
Standard Charges for Core Facility
Flow cytometry analyis – FACScan,FACSCalibur, LSRII
Operator Assisted: $40/hr
Independent user: $20/hr
Cell sorting – MoFlo, FACSAria
Set-up fee: $13
Hourly rate: $92/hr
Corporate users
Flow Cytometry Analysis: $50/hr
Flow Cytometry Sorting: $200/hr and Setup Fee: $25
What is Cytometry? and Learning Diva Software:
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/
Fluorescence Spectrum Viewers:
http://probes.invitrogen.com/resources/spectraviewer/
Fluorochrome information:
http://www.zeiss.com/C12567BE0045ACF1/Contents-Frame/CF19CAEC78522802C1256AB0005242E9
Our Beckman Coulter Moflo XDP sorter purchased in 1998 is equipped with an Enterprise II 621 laser, a Coherent Cube 640 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-CY7.
A BD Biosciences FACSAria IIu 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 hematopoietic progenitors or B cell subsets from tonsils. Finally, the 407 nm laser is appropriate for FRET experiments, exciting Quantum Dots and for using Hoechst 33342 to study Side Population. The FACSAria also has an Aerosol Containment Device to ensure that any aerosolized particles are contained properly and making it safer to run human tissue samples. The LSRII analyzer is equipped with 405 nm, 488 nm, 561 nm and 633 nm lasers. This digital instrument is a good companion to the FACSAria and is capable of detecting up to 16 parameters.
A BD Biosciences FACScan and FACSCalibur cytometers are capable of three and four color fluorescence analyses, respectively.
To assist researchers with analysis, the Flow Lab has a MacMini with Cellquest Pro and FlowJo and a PC with Diva andSummit. We also have three dongles for the Diva, FlowJo and Cellquest Pro that can be loaned to users. We also provide a transfer file on the sanserver and have CDs and DVDs available to make copies of data files. Although we make backups of the computers, all users should frequently back-up their data.
Detailed Equipment Specifications
