The Flow Cytometry Shared Resource (FCSR) provides a range of instrumentation and expertise for cell sorting and analysis that can support experimental needs related to single cells, cell line production, biosensor development, and dye analysis, allowing researchers to develop new tools and answer questions about the nature and function of cells. The FCSR has expertise in flow cytometry experimental design, operation of instruments, distribution of raw data and sort reports, interpretation of results, and preparation of graphics for presentation and publication.
Flow Cytometry Instruments
The Cytoflex S is a cell analysis flow cytometer equipped with four excitation lasers (405nm Violet, 488nm Blue, 561nm Yellow-Green, and 638nm Red), and 13 fluorescence channel detectors. Light scatter can be detected off the Violet laser for small particle/exosome analysis. Samples can be loaded from traditional 12x75mm flow tubes as well as from 96 well plates. This instrument is used for analysis of fluorescence signal but not for sorting.
The Sony SH800 is a sorting flow cytometer capable of sorting two populations simultaneously into tubes and well plates of various sizes (6, 12, 24, 48, 96, and 384). Using three nozzle sizes (70um, 100um and 130um), cells can be deposited as single cell isolates into wells, or a collection of cells in wells or tubes. The sorter is equipped with four colinear lasers (405nm, 488nm, 561nm, and 638nm; single interrogation site) and has 6 fluorescence detection channels.
BD FACSAria SORP
The FACSAria is a sorting flow cytometer capable of sorting up to four populations simultaneously into tubes, well plates of various sizes (6, 24, 48, 96 or 384), and onto glass slides. Using four nozzle sizes (70um, 85um, 100um or 130um), cells can be deposited as single cell isolates into wells, or a collection of cells in wells or tubes. The sorter is equipped with five spatially separated lasers (405nm, 445nm, 488nm, 561nm, and 637nm; 5 interrogation sites) and has 16 fluorescence detection channels. This sorter is contained within a Baker BSC for sorting BSL-2 samples.
Bigfoot 9 Laser Spectral Sorter
The Bigfoot is a sorting flow cytometer capable of sorting up to six populations simultaneously into tubes, well plates of various sizes (6, 12, 24, 48, 96, 384, 1536), and other custom devices, including 10X Genomics chips. Using five nozzle sizes (50um, 70um, 100um, 120um or 150um), cells can be deposited as single cell isolates into wells, or as a collection of cells in wells or tubes. Plate sorting is done with multiple streams (4 streams on 96 well plates and 8 streams on 384 well plates), increasing the collection speed by greater than 10x over single stream deposition. This instrument is equipped with nine lasers (UV 349nm, 405nm, 445nm, 488nm, 532nm, 561nm, 594nm, 640nm, 785nm) and has 60 fluorescence channel detectors. Seven of the lasers are spatially separated and two are collinear, equating to seven interrogation sites. In addition, this sorter has an integrated BSC, an automated sample loader for six independent samples, an integrated wash station, and on-board calibration beads. As a result, the Bigfoot cell sorter can analyze a single cytometry sample that contains more than 40 different colors and captures the bulk of the emission spectra of each fluorophore. This allows for fluorescent signal separation, which increases the number of fluorochromes possible per panel and the resolution of close emission fluorophores.