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Flow Cytometry Technologies

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
  • 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.

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 labelled 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.
FACS Aria Fusion, 11 color, flow cytometric cell sorter
FACS Aria II, 10 color, flow cytometric cell sorter
FACS Aria IIu, 17 color, flow cytometric cell sorter
FACS Canto-II, 8 color, Flow Cytometric Analyzer
LSR Fortessa HTS, 19 color, high throughput, flow cytometric analyzer
LSR II, 12 color, flow cytometric analyzer
CellCelector, ALS
Helios A CyTOF System
Helios A CyTOF System
Hyperion Imaging System
Invitrogen Countess cell counter
Zeiss fluorescence microscope