Cell Sorting Instrumentation

Our BSL2+ room C315 houses the two BD FACSAria cell sorter instruments.

The BD FACSAria I was upgraded in 2010 and comes equipped with 12 fluorescent PMTs over four lasers (405 nm violet, 488 nm blue, 561 nm yellow-green, and 640 nm red). This configuration is similar to the LSRII B analyzer, but with less options off the violet laser. The instrument operates with a 100 micron nozzle/20 PSI or 70 micron nozzle/35 PSI configuration. Detailed specifications can be found in the menu on the right. The Aria I is placed in a Bio-Bubble negative-pressure biosafety cabinet that provides the BSL2+ environment to protect the operator.

The BD FACSAria II is equipped with four lasers (355 nm UV, 405 nm violet, 488 nm blue, and 633 nm red) and has 13 fluorescent detectors. This configuration is similar to the Fortessa B analyzer, but with less options off the violet laser. Current options for configuration on the Aria II are 100/20; 85/45; 85/30; 70/35, and 130/10 (nozzle size/PSI). Detailed specifications can be found in the menu on the right.

Both instruments have the capacity to do one- to four-way sorts into a variety of tube styles, such as 12x75 mm tubes of any material, Eppendorfs, PCR tubes, and 15-ml conical tubes (2-way only). They can also sort into plates (6, 12, 24, 48, 96, and 384 wells) and onto slides.

Our cell sorters undergo a stringent routine maintenance and cleaning protocol to warrant aseptic sorting conditions at all times. We use sterile-filtered HEPES-buffered saline with EDTA as sheath fluid.

For magnetic cell sorting, our facility houses a Miltenyi Classic AutoMACS. This instrument is located in a biosafety cabinet in Rm C322, and does not require operator assistance. Columns, clean bottles, and cleaning solution are provided. The user is responsible for running buffer and beads. Daily usage charges apply. Training is available through Kitty de Jong x8416.