The Flow Cytometry Shared Resource is located in a 1,243 square-foot, open-access/limited-access facility: RMSB. 7147. The shared resource provides the following instruments:

• BD FACS Canto-II, 8-color, flow cytometric analyzer 
• BD LSR-II, 12-color, flow cytometric analyzer
• BD LSR-Fortessa-HTS, 19-color, high-throughput, flow cytometric analyzer
• Cytek Aurora, 64 fluorescence channels, spectral analyser, with auto-loader. 
• BD FACS Aria Fusion, 15-color, flow cytometric cell sorter, with integrated BSL-2 bio-containment
• BD FACS Aria-IIu, 17-color, flow cytometric cell sorter, housed in a Baker BioProtect-II, BSL-2 bio-safety cabinet
• BD FACS Aria-II, 10 color, flow cytometric BSL-1 cell sorter
• Fluidigm Helios CyTOF mass cytometer
• Fluidigm Hyperion imager
• CellCelector automated, bio-contained, cell transfer robot 
• Zeiss fluorescence microscope
• Invitrogen Countess cell counter 

Access to the FCSR’s instruments 

Individual users can be trained independently to operate the Canto-II, LSR-II and LSR-Fortessa-HTS flow cytometric analyzers. Cell sorting is performed with the three FACS Arias, by the facility’s operators only. Sorting is available by prior request for after-hours and weekend operation. Mass cytometry also is performed only by the facility’s staff. 

Instruments

Analysis 

• FACS Canto-II Analyzer
  Three-laser, fully digital FACS Canto-II analyzer, used for up to eight-color (ten-parameter) analysis of cell samples. Runs DiVa-8 software.
  
  
• LSR-II Analyzer
  Sophisticated, analytical instrument, used for complex analysis of cell samples. Recently upgraded, now with four lasers, for 12-color analysis. DiVa-8 software.
  
  
• LSR-Fortessa-HTS analyzer
  Sophisticated, analytical instrument, used for complex analysis of cell samples. Five lasers, for 19-color analysis. DiVa-8 software. HTS high-throughput automated sample loader, for rapid analysis of stained cells directly from culture plates.
  
  
• Cytek Aurora, Spectral analyzer
  Five laser, 64 fluorescence detection channels and 3 scatter channels. SpectroFlo software. Automatic Micro-Sampling System (loader), for rapid analysis of samples directly from 96-well plates. For each fluorochrome, a full fluorescence emission spectrum is generated and stored as a precise spectral vector “fingerprint” for that specific marker. These spectra are presented as ribbon plots and added to a spectral reference library. This novel technology allows the rapid simultaneous analysis of reagent combinations which previously were unresolvable, because of spectral interactions. 
  

Sorting

FACS Aria Cell Sorters – Three FACS Aria cell sorters, used for complex analysis and sorting of cell samples. They incorporate a fixed-alignment flow cell, for enhanced sensitivity of fluorescent detection and ease of optical alignment. All have “Whisper” aerosol evacuation systems and DiVa-8 software. 

• BD FACS Aria Fusion - Custom-built (SORP), sophisticated, high-speed cell sorter, with sample temperature control and an Automatic Cell Deposition Unit (ACDU), for sorting on to culture plates and for single cell flow-cloning. Four lasers, for up to 15-color simultaneous fluorescence analysis. Integrated bio-containment, for sorting of hazardous BSL-2 samples. 
• BD FACS SORP Aria-IIu - SORP high-speed cell sorter, with an ACDU and temperature control. Five lasers, for 17-color analysis. Baker BioProtect-II bio-safety cabinet, for sorting of BSL-2 samples. 
• BD FACS SORP Aria-II – SORP high-speed cell sorter, with four lasers, for 10-color simultaneous analysis. BSL-1 samples only.

Mass cytometry

• Fluidigm CyTOF Helios mass cytometer - 30+ parameter cytometry of labeled cells in suspension, using time-of-flight mass spectrometry of target-specific heavy metal-tagged antibodies, vapourised in a plasma. 
• Fluidigm Hyperion imager - Simultaneous detection of over 30 protein markers with sub-cellular resolution in solid tissue sections, from a single scan. An ablating laser beam scans at one square micron pixel resolution and then analyzes the plume of vaporized metal tags by CyTOF mass cytometry.

Other

• Zeiss fluorescence microscope – Used for the examination of fluorescent cell samples, before and/or after cell sorting.
• Invitrogen Countess cell counter – Uses 10ul. of sample for a total cell count, plus cell viability analysis using Trypan Blue dye. 
• Data analysis software – Site licenses for the FlowJo, FCS Express and Cytobank packages, with seats on each available for annual fees. Astrolabe account, to access fee-for-service high parameter (eg. CyTOF) data analysis.