Flow Cytometry

Contact: Betty Anne McBey or Dr. Dorothee Bienzle or Dr. Byram Bridle

Betty Anne McBey (bmcbey@uoguelph.ca, 519-824-4120 ext. 54758 or 54733) operates the FACSAria IIu and trains individuals to use the FACSCanto II and Accuri C6.


The Flow Cytometry Core Facility in the Department of Pathobiology has the following equipment available for on-campus and 

off-campus users. We welcome inquiries about any kind of cytometry experiment - our previous experience includes sorting and analysis of equine stem cells, chloroplasts and other plant components, porcine spermatozoa, transfected cells, lymphocytes from many species, platelets, and more.


NOTE: It takes planning and optimization to set up parameters for cell sorting on the FACSAria (fluorochromes, cell type, size, complexity, expected proportion of positive cells, downstream application for cell culture, RNA, DNA, or protein extraction, etc.). We respond to any inquiries within a day, but it may take several test samples to set up proper sorting. 



FACSAria IIu Cell Sorting System (FACSAria manual)























Characteristics
  • High-speed cell sorting
  • 3 air-cooled lasers (blue: 488, red: 633, and violet: 407 nm)
  • Digital acquisition rates of up to 70,000 events/second
  • Multicolour analysis (up to 9 colours)
  • Two- and four-way bulk sorting into tubes and 96-well plates
  • Our experience: 85-99% purity of sorted avian, bovine, and feline lymphocytes
  •  Optical filters to detect fluorophores in the following nine channels:

Blue (488 nm) laser:

530/30 nm (e.g. FITC, Alexa Fluor 488, eGFP, CFSE)

585/42 nm (e.g. PE, propidium iodide)

616/23 nm (e.g. PE-Texas Red)

695/40 nm (e.g. PerCP, PerCP-Cy5.5, 7-AAD)

780/60 nm (e.g. PE-Cy7)

Red (635 nm) laser:

660/20 nm (e.g. APC, Alexa Fluor-647)

780/60 nm (e.g. APC-Cy7)

Violet (405 nm) laser:

450/40 nm (e.g. Pacific Blue, violet proliferation dye 450, Horizon V450, brilliant violet 421)

530/30 nm (e.g. AmCyan)

User fees (sorting):

Departmental users: $70/hr for first 3 hrs, $40/hr for additional hours.

External: users: $110/hr

User fees (analysis only):

Departmental users: $40/hr

External users: $70/hr


FACSCanto II (FACSCanto manual)