Instruments
Cell Sorters
The sorters in our Facility make use of electrostatic deflection of droplets. Fluid containing particles passes through a nozzle and a stream emerges into the air. The fluid stream breaks up into droplets- with the aim being the generation of droplets big enough to contain one cell in each drop. To do this, the stream is vibrated at a specific frequency and pressure to separate it into droplets. These droplets are then charged or not depending upon the fluorescent profile of the cell(s) within. The drops pass through an electric field allowing electrostatic or droplet deflection which sends the charged drops of interest into the collection vessel. Unsorted droplets go into the waste.
BD Aria III | Instrument Configuration
The BD FACSAria III cell sorter is the high-speed benchtop digital flow cytometer. It is equipped with five spatially separated air-cooled lasers - 488 nm, 633 nm, 561 nm, 375 nm, and 405 nm. The near UV (375 nm) laser uses the same path as the violet (405 nm) laser and cannot be used simultaneously.
Most cell types can be accommodated with the use of 130, 100, 85, or 70-micron nozzles. Sorting can be performed into two or four tubes with a speed of up to 35,000 events/sec. The Automated Cell Deposition Unit (ACDU) makes it possible to sort individual cells into multi-well plates (up to 96 well), PCR tubes, or microscope slides. The instrument is also equipped with Aerosol Management Option (AMO), a device that traps aerosolized particles. The system operates on a PC-based Windows platform using BD FACSDiva software.
Location: Room 667 Heritage Medical Research Centre (HMRC).
Sony MA900 (12-color)
The MA900 system includes a patented microfluidic chip-based design, comprehensive fluidic controls, and advanced automation. It is equipped with 4 excitation lasers - 488 nm, 405 nm, 561 nm, and 638 nm - on two beam spots. Free-form PMTs enable the detection of fluorescence signals from each beam spot, allowing the detection of 12 fluorescence parameters and 2 scatter parameters. This sorter is available for user training and operation.
The Sony MA900 is enclosed by the BioProtect IV ClassII Type A2 biosafety cabinet from Baker Company. The cabinet provides HEPA filtration and airflow control to maximize the protection of the operators from potentially hazardous samples.
The MA900 has microfluidic chips available for 100 or 70-micron sorting. Sorting can be performed into two or four tubes or into multi-well plates (up to 384 well) or PCR tubes. Index sorting is available in multi-well plates. Index sorting software records the X and Y coordinates of each event sorted into a multiwell device. This software enables you to track the scatter and fluorescence intensity of individual cells sorted in each well. The instrument is also equipped with Aerosol Management Option (AMO), a device that traps aerosolized particles.
Location: Room 667 Heritage Medical Research Centre (HMRC).
Cell Analyzers
Our Flow Cytometry analysers are based on differential pressure and are designed to analyze cells at a high rate (up to ca 10,000 - 40,000 cells (events) per second on the perfect single cell sample).Several parameters can be measured on individual cells within a few minutes. This is the power of flow cytometry. The properties measured include a particle's relative size, granularity, and relative fluorescence intensity (please see the individual machines for the number of fluorochromes they can analysis in one cell/particle). Any suspended particle or cell from 0.2-70 micrometers in size is suitable for analysis. Larger or smaller particles require specialized cytometers/conditions.
Attune NxT (14 color, tubes) | Instrument Configuration
The Attune NxT is a benchtop digital flow cytometer that uses acoustic focusing to allow for extremely high event rates (35,000 events/ second). Equipped with 4 spatially separated lasers (405nm, 488 nm, 561nm, and 637 nm) it is capable of analyzing forward and side scatter and 14 fluorochromes with excellent sensitivity for detection of dim events. Digital electronics allow automated setup and compensation calculation, as well as post-acquisition compensation.
Location: Room 667 Heritage Medical Research Centre (HMRC).
BD LSR Fortessa-SORP (BSL1) | Instrument Configuration
The LSR-Fortessa SORP is a benchtop digital flow cytometer. Equipped with 5 air-cooled spatially separated lasers (375nm, 405nm, 561nm, 488 nm and 640 nm) it is capable of analyzing forward and side scatter and 18 fluorochromes with excellent sensitivity for detection of dim events.
Location: Room 667 Heritage Medical Research Centre (HMRC).
Cytek Aurora
The Cytek Aurora is a digital spectral flow cytometer. It is equipped with 5 lasers (355 nm, 405 nm, 488 nm, 561 nm, and 640 nm) and a 65 detector array. The Aurora system delivers high-resolution data at the single-cell level to resolve the most challenging cell populations, such as cells with high autofluorescence or low levels of expression of key biomarkers, regardless of assay complexity. It is a high-parameter flow cytometer, capable of analyzing 40+ parameters.
Location: Room 667 Heritage Medical Research Centre (HMRC).
Cytek Amnis ImageStream mkII (BSL1)
The Cytek Amnis ImageStream mkII is a bench-top imaging flow cytometer. It is equipped with 3 lasers (488 nm, 642 nm and 405 nm) and is capable of analyzing 9 fluorochromes, side scatter, and bright field images. It is equipped with 20X, 40X, and 60X magnification, high gain mode, and an autosampler. Please refer to the table here (ImageStream lasers) for a list of suitable fluorochromes- a table is pasted below for a quick reference. This system operates on a PC-based Windows platform using INSPIRE software.
Location: Room 667 Heritage Medical Research Centre (HMRC).
Cytek Northern Lights 3000
The Cytek Northern Lights is a digital spectral flow cytometer. It is equipped with 3 lasers (405 nm, 488 nm, and 640 nm) and a full spectrum profiling (FSP) detection system. The Northern Lights system delivers high-resolution data at the single-cell level to resolve the most challenging cell populations, such as cells with high autofluorescence or low levels of expression of key biomarkers, regardless of assay complexity. It is a high-parameter flow cytometer, capable of analyzing 20+ parameters.
Location: Room 6-127 Katz Group Centre for Pharmacy & Health Research (KATZ).
BD Fortessa X-20 (BSL2) | Instrument Configuration
The LSR-Fortessa X20 is a benchtop digital flow cytometer. Equipped with 5 air-cooled spatially separated lasers (375 nm, 405 nm, 488 nm, 561 nm, and 633 nm) it is capable of analyzing forward and side scatter and 18 fluorochromes, with excellent sensitivity for detection of dim events. This unit is also contained within a BioBubble to permit analysis of BSL2 samples. The system operates on a PC-based Windows 7 platform using BD FACSDiva software. Digital electronics allow automated setup and compensation calculation, as well as post-acquisition compensation.
Location: Room 6-125 Katz Group Centre for Pharmacy & Health Research (KATZ).
Small Particle Instruments
NanoView Biosciences ExoView R200
The NanoView Biosciences ExoView R200 allows for fully automated, purification-free detection of exosomes, extracellular vesicles (EVs), and viruses. Using a chip-based technology, this instrument can provide high-resolution size distribution of EVs between 50-200nm, phenotyping and co-localization of markers in EV membranes, detection of EV cargo proteins, EV count, and viral size and titer
Location: Room 667 Heritage Medical Research Centre (HMRC).
Beckman Coulter CytoFLEX-S | Instrument Configuration
This instrument has excellent sensitivity of detection using avalanche photodiodes, violet SSC for small particle detection, and a 96-well plate loader for high throughput applications. NOTE: This instrument will be limited to small particle work. No cells can be run on this platform.
Location: Room 667 Heritage Medical Research Centre (HMRC).