Cytometry and Imaging Shared Resource major research instruments include:
| Type | Instrument |
|---|---|
| Sorters | Cytek Aurora CS |
| BD FACS Aria Fusion | |
| BC CytoFLEX SRT | |
| Analyzers | BD FACSymphony A5 SE |
| Cytek Aurora | |
| BD LSR Fortessa HTS | |
| BD FACS Canto-II | |
| CyTOF | Standard Biotools Helios |
| Standard Biotools Hyperion | |
| Confocal Microscope | Zeiss LSM 980 w/AiryScan2 |
- BD FACS Canto-II: cell analyzer with 8 fluorescent parameters and 2 light-scatter parameters.
- BD LSR-Fortessa-HTS: cell analyzer with 18 fluorescent parameters and 2 light-scatter parameters.
- BD FACSymphony A5 SE: cell analyzer with 30+ fluorescent parameters and 2 light-scatter parameters using both conventional compensation and spectral unmixing platforms.
- Cytek Aurora: cell analyzer with 40+ fluorescent parameters and 3 light-scatter parameters using the spectral unmixing platform.
- BD FACS Aria Fusion: BSL-2 cell sorter with 15 fluorescent parameters and 2 light-scatter parameters.
- Cytek Aurora CS: BSL-2 cell sorter with 40+ fluorescent parameters and 3 light-scatter parameters using the spectral unmixing platform.
- Beckman Coulter CytoFLEX SRT: BSL-2 cell sorter with 15 fluorescent parameters and 2 light-scatter parameters available for self-operated cell sorting 24/7/365.
- Standard BioTools Helios CyTOF Mass Cytometer.
- Standard BioTools Hyperion Imaging Mass Cytometer.
- Zeiss LSM 980 laser scanning confocal microscope with Airyscan2
Use of the CISR’s instruments
The facility offers virtual and hands-on training sessions to individual users for self-operated use on all the cell analyzers (FACS Canto-II, LSR-Fortessa-HTS, FACSymphony A5 SE and Cytek Aurora) and the confocal microscope (Zeiss LSM980), with operator assistance available upon request for all users (trained and untrained). FACS Aria Fusion and the Cytek Aurora CS cell sorters are operator-assisted, with the Beckman Coulter CytoFLEX SRT cell sorter available as an operator-assisted or self-operated instrument. Self-operated cell sorting on the CytoFLEX SRT requires comprehensive training offered by the CISR. The CyTOF Helios Mass Cytometer and Hyperion Imaging Mass Cytometer are operator-assisted. Experience and trained internal users can request 24/7/365 access to all self-operated instruments, which will be granted after demonstrating proficiency in proper instrument operation and shutdown.
Instruments
Analysis
- BD FACS Canto-II Analyzer – The FACS Canto-II is a 3-laser flow cytometric cell analyzer with 8 fluorescent parameters and 2 light-scatter parameters. Operates using FACSDiva 9 acquisition software on a PC workstation.
- BD LSR-Fortessa-HTS Analyzer – The LSR-Fortessa-HTS is a 5-laser flow cytometric cell analyzer with 18 fluorescent parameters and 2 light-scatter parameters.Equipped with a high-throughput automated sample loader for rapid analysis of stained cells directly from a 96-well plate. Operates using FACSDiva 9 acquisition software on a PC workstation.
- Cytek Aurora, Spectral Analyzer – The Aurora is a 5-laser flow cytometric cell analyzer with 64 fluorescent detectors and 3 light-scatter parameters capable of analyzing up to 40+ fluorescent parameters using the spectral unmixing platform.Equipped with an Automatic Micro-Sampling System for rapid analysis of samples directly from a 96-well plate.Each fluorochrome has a unique spectral signature across the full fluorescence emission spectrum. As a spectral analyzer, the Aurora can separate reagent combinations that were previously unresolvable due to spectral interactions. Operates using SpectroFlo acquisition software on a PC workstation.
- BD FACSymphony A5 SE Analyzer – The FACSymphony A5 SE is a 5-laser flow cytometry cell analyzer with 48 fluorescent detectors and 2 light-scatter parameters capable of analyzing 30+ fluorescent parameters using both conventional compensation and spectral unmixing platforms. The instrument features an optimized filter set to collect the full spectrum of emitted light allowing for simultaneous analysis of fluorochromes with similar spectral signatures. Operates using FACSDiva 9 acquisition software on a PC workstation.
Cell Sorting
- Becton Dickinson (BD) FACS Aria Fusion – The FACS Aria Fusion is a 4-laser high-speed cell sorter with 15 fluorescent parameters and 2 scatter parameters.Equipped with sample temperature control and an Automatic Cell Deposition Unit (ACDU), for sorting directly into 96-well plates for tissue culture and single cell cloning. The instrument includes an integrated bio-safety cabinet for processing hazardous samples up to BSL-2+ classification.
- Cytek Aurora CS – The Cytek Aurora CS is a 5-laser spectral analysis cell sorter capable of analyzing 40+ fluorescent parameters and 3 scatter parameters. The instrument is in a bio-safety cabinet for sorting BSL-2+ samples. Equipped with sample temperature control for input and output, the Aurora CS can sort up to 6-ways simultaneously or into 96 and 384-well plates. As a spectral analysis instrument, autofluorescence can be subtracted to allow for better identification markers in samples with high fluorescence backgrounds.
- Beckman Coulter CytoFLEX SRT – The CytoFLEX SRT is a 4-laser cell sorter with 15 fluorescent parameters and 2 scatter parameters.Equipped with sample temperature control and a CyClone movement system, for sorting into tubes, slides or multiwell plates.Housed in a bio-safety cabinet for processing of hazardous samples up to BSL-2+ classification. Operates using CytExpert SRT acquisition software on a PC workstation, the CytoFLEX is designed for ease-of-operation.
Mass Cytometry
Standard Biotools Mass Cytometry
- CyTOF Helios (Suspension Mass Cytometry) – The CyTOF Helios Mass Cytometer analyzes single cell suspensions labeled with antibodies conjugated to heavy metal isotopes (50+ distinct metal isotopes).The cells are nebulized and sent through argon plasma, which results in an ion cloud.This ion cloud is processed, filtered and then detected in a time-of-flight mass spectrometer. Operates using CyTOF Software 7.0 on a PC workstation.
- CyTOF Hyperion (Imaging Mass Cytometry) – The Hyperion Imaging Mass Cytometer analyzes FFPE or frozen solid tissue sections or fixed cells that are adhered to a slide and labeled with up to 50+ antibodies conjugated to heavy metal isotopes.A UV laser ablates the tissue of interest in 1 micron square increments (“pixels”) and ionized and detected in a time-of-flight mass spectrometer.Multiplexed images are generated at subcellular resolution. Operates using CyTOF Software 7.0 on a PC workstation.
Confocal Microscopy
- Zeiss LSM 980 Confocal Microscope with Airyscan 2 - provides confocal optical sectioning of fixed or live samples, with full environmental control (CO2 and temperature) for time-lapse imaging. This system utilizes an AI-mediated sample finder and can accommodate carriers, including slides, chambered slides, multi-well plates, and Petri dishes for multi-position imaging. It is also capable of wide-field imaging using an LED light source, transmitted light imaging, spectral unmixing, tile-scanning, and has an Airyscan 2 super resolution module for 120 nm resolution (down to 90 nm resolution with deconvolution).
Other
- Data analysis software – Site licenses for FlowJo, FCS Express, TerraFlow and Visiopharm available with annual fees per license or per use.Visiopharm AI-assisted image analysis for Hyperion imaging mass cytometry data. Data analysis is available for certain applications on a fee-for-service basis.
FACS Aria Fusion, 15 color, flow cytometric cell sorter
FACS Canto-II, 8 color, Flow Cytometric Analyzer
LSR Fortessa HTS, 19 color, high throughput, flow cytometric analyzer
Helios A CyTOF System
Helios A CyTOF System
Hyperion Imaging System