Cytometry and Antibody Technology

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CAT Facility description:

Cytometry and Antibody Technologies Facility (CAT):  https://voices.uchicago.edu/ucflow/   This facility is directed by Position vacant. The main facility is located in the Biological Sciences Learning Center and has a satellite facility in the Albert Merritt Billings Hospital.  The CAT Facility serves the faculty by providing access to state-of-the-art technology in quantitative analytical approaches to measure molecular and cellular function. The facility is designed to meet the widespread needs for specialized cytometric analysis and continues to respond to the demand for new and improved technology. The CAT facility supports a Helios mass cytometer, which allows for the detection of 40+ parameters on a cell-by-cell basis. It also supports two 5-laser (355nm, 405nm, 488nm, 561nm and 640nm) Cytek Aurora flow spectral analyzer which allows for the measurement on 40+ fluorophores on every single cell. Currently the main facility employs 4 standard cytometers, including a ThermoFisher AttuneNxT with 4 lasers (405nm, 488nm, 561nm and 640nm) and 14 detectors, two custom 4 laser (405nm, 488nm, 561nm, and 640nm) / 15 parameter detectors BD LSR-Fortessa (one equipped with a 96 well HTS unit), and a custom-built BD LSRII equipped with 4 lasers (405nm, 488nm, 561nm, and 640nm) and 12 parameter detectors. In addition, the CAT facility has a Bigfoot Spectral cell sorter equipped with 5 lasers and 55 detectors. It also hosts a BD FACSAria high-speed cell sorters equipped with 4 lasers (405nm, 488nm, 561nm and 640nm) and 15 parameter detectors, as well as a BDFACS Aria Fusion cell sorter equipped with 5 lasers (UV, 405nm, 488nm, 561nm, and 640nm) and 18 parameter detectors. This last instrument, incased in a BSL2 laminar flow hood, is dedicated solely for users processing human patient samples in a translational research setting. All of the cell sorters are equipped with 96-well plate robotic arm attachments, various nozzle tip sizes (70um – 130um) and temperature control (4C – 37C). The CAT Facility also hosts a MACSQuant Tyto cell sorter equipped with 3 lasers (488nm, 405nm, and 638nm) and 8 fluorescence detectors which process cells within a closed cartridge, offering full biosafety and protection against microbial contamination. The satellite facility has an Agilent Quanteon equipped with 4 lasers (405nm, 488nm, 561nm and 637nm) capable of measuring up to 16 parameters. Additionally, the facility has an imaging cytometer, the Luminex ImageStreamX MkII, which has 4-lasers (405nm, 488nm, 561nm, 640nm) and two 6-channel CCD cameras to collect up to 10-color fluorescence image data from thousands of cells per second. The facility also has a dedicated Luminex 200 bead analyzer from Millipore and a Quickplex SQ120 from Meso Scale Discovery to perform multiplex cytokine assays.  Trained users are provided 24-hour access on any of these instruments with the exception of the Helios mass cytometer, which is operated by the staff of the facility.

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Acknowledging the Facility or the staff and listing the Core equipment you used in the Materials and Methods section helps us demonstrate how much our Core contributes to the University community and track the impact of each piece of equipment on research infrastructure. Using the Facility RRID helps us track publications where we are acknowledged. Facility RRID: SCR_017760

General Acknowledgments:

[Technique/technology] was performed at the Cytometry and Antibody Technology Facility at University of Chicago, which receives financial support from the Cancer Center Support Grant (P30CA014599). RRID: SCR_017760

We thank The University of Chicago Cytometry and Antibody Technology Facility, especially [staff name], for their assistance with [technique/technology]. RRID: SCR_017760

 

Instrument-Specific Information:

Benchtop Analyzers

  • Fortessa 4-15: Samples were acquired on a LSRFortessa running FACSDiva version 8.0.2 (Special Order Research Product, BD Biosciences), equipped with 15 detectors and 405nm, 488nm, 561nm, and 640nm lasers.
  • Fortessa 4-15 HTS: Samples were acquired on a LSRFortessa running FACSDiva version 8.0.2 (Special Order Research Product, BD Biosciences), equipped with 15 detectors and 405nm, 488nm, 561nm, and 640nm lasers.
  • Fortessa X-20 (retired in June 2021): Samples were acquired on a LSRFortessa X-20 running FACSDiva version 8.0.2 (Special Order Research Product, BD Biosciences), equipped with 18 detectors and 355nm, 405nm, 488nm, 561nm, and 640nm lasers.
  • LSRII 4-12: Samples were acquired on a LSR II running FACSDiva version 8.0.2 (Special Order Research Product, BD Biosciences), equipped with 12 detectors and 405nm, 488nm, 561nm, and 640nm lasers.
  • Attune NxT: Samples were acquired on an Attune NxT running Attune Cytometric (Thermo Fisher Scientific), equipped with 14 detectors and 405nm, 488nm, 561nm, and 640nm lasers.
  • Aurora Blue/Green/PME/Red: Samples were acquired on an Aurora running SpectroFlo version *see below* (Cytek Biosciences) equipped with 64 detectors and 355nm, 405nm, 488nm, 561nm, and 640nm lasers.
    • Software versions:
      • Current (Red, installed Nov 15, 2021): 3.0.1
      • Current (PME): 2.2.0.4
      • Previous: 3.0 (Red, Sept 30-Nov 14, 2021)
      • Previous: 2.2.0.4 (was released around December 2020)
      • Previous: Version 2.2.0.3 was installed around the middle of September 2020
      • Previous: Version 2.2 was installed around the end of January 2020

Cell Sorters

  • Aria Fusion 5-18: Samples were sorted on a FACSAria Fusion running FACSDiva (Special Order Research Product, BD Biosciences), equipped with 18 detectors and 355nm, 405nm, 488nm, 561nm, and 640nm lasers.
  • AriaII 4-15: Samples were sorted on a FACSAria II running FACSDiva (Special Order Research Product, BD Biosciences), equipped with 15 detectors and 405nm, 488nm, 561nm, and 640nm lasers.
  • AriaIIIu (retired in 2021): Samples were sorted on a FACSAria IIIu running FACSDiva (BD Biosciences), equipped with 15 detectors and 405nm, 488nm, 561nm, and 640nm lasers.
  • AriaIII 3-14 (rebuilt November 2021): Samples were sorted on a FACSAria III running FACSDiva (BD Biosciences), equipped with 14 detectors and 405nm, 488nm, and 640nm lasers.
  • Tyto: Samples were sorted on a MACSQuant Tyto (Miltenyi Biotec), equipped with 8 detectors and 405nm, 488nm, and 635nm lasers.

Other

  • ImageStream: Samples were acquired on an Image Stream MarkII (Luminex), equipped with 2 cameras divided in 12 channels and 405nm, 488nm, 561nm, and 640nm lasers.
  • Luminex: Samples were acquired on a Luminex 200 (Luminex)
  • MSD Quickplex: Samples were acquired on a Quickplex SQ120 (Meso Scale Discovery)