FACSAria IIu is a benchtop High Speed Cell Sorter produced by Becton Dickinson, with 3 lasers and 9 colors, upgradable to 13 colors. Optical quality control and calibration are performed daily. Typical fluorescent labels detected are Pacific Blue, CFP, FITC, Alexa488, GFP, CFSE, YFP, PE, PE-TexasRed, PerCP, PerCP-Cy5.5, PE-Cy5, PI, 7-AAD, PE-Cy7, APC, Cy5, APC-Cy7, etc). For more information, check the following topics:
FACSAria IIu is equipped with 3 different low-power lasers that can be used simultaneously. A 13mW solid-state blue (488nm) laser, from which SSC and another 5 colors can be detected (upgradable to another 2); an air-cooled 11mW HeNe red (633nm) laser, from which 2 colors can be detected (upgradable with an extra color); and a solid-state 10mW violet (407nm) laser, from which 2 colors can be detected (also upgradable with an extra color). Instead of calling the detection channels generically FL1-FL9, BD named each channel after the most typical fluorochrome it detects (see picture below). Note that a band-pass filter designated as 530/30 indicates that it will detect light in the a 30nm range centered at 530nm, i.e., 515-545nm. LP stands for Long Pass filter, and indicates that it will detect light above the mentioned wavelength. A list of typical fluorescent labels detected in each channel is also displayed on the right.
After switching off the instrument, wipe every surface that has been exposed to sheath fluid, to prevent salt build up.
Cleaning the Waste Tank:
FACSAria is connected to a PC loaded with BD FACSDiva Software for machine interface and data acquisition. Whether you're sorting or acquiring, FACSDiva Experiments are saved within the actual software (in one single database file, containing all the other saved experiments). To avoid reaching the limit size of this database file ~16GB, all Diva Experiments should be exported, and after confirming there is no data loss, they should be deleted from within the software. The way the data is exported, depends on where the data is to be analyzed. If you wish to maintain the same plots and gates drawn in the FACSDiva software, you must export the actual Experiment and then you may import it back in any offline FACSDiva software. To analyze the data, for instance, with Flowjo, data must be exported as FCS files (the "Flow Cytometry Standard"). Depending on the software and its version, you may have to choose FCS 2.0 or FCS 3.0.
Once data has been exported to the hard drive transfer it by saving it to a pen-drive or transferring it to your directory in xserve02. To transfer data to xserve02, follow these steps:
IMPORTANT: After you have checked there is no data loss in your exported experiment, delete the Experiment from the FACSDiva Software. FACSDiva software is prone to crashes and "mysterious" data losses have been reported by the flow cytometry community. The first thing to do after any flow analysis or sorting, is to save your own data.
Though we don't have any real custom changes in the FACSAria, we do have some optional components: