Instrumentation
The Flow Cytometry Facility is equipped with one high-speed cell sorter (FACSAria Fusion), one imaging cytometer (ImageStream X Mk II), one full-spectrum analyser (Aurora) and four conventional cytometers (LSRFortessa, FACSCanto II, Accuri C6 and Attune NxT). An in vivo fluorescence & luminescence imager (Biospace Lab PhotonIMAGER RT) is housed in the Comparative Medicine Unit.
FACSAria Fusion Cell Sorter:
The FACSAria Fusion (BD Biosciences) combines a top of the range high speed cell sorter with best-in-class biosafety. Housed in a Class II Type A2 biosafety cabinet, the sorter performs with unrivalled sensitivity and resolution. With 4 nozzle sizes (70, 85, 100, & 130 µm), 4 lasers (405, 488, 561, & 640 nm) and 16 fluorescence parameters, this provides a vast array of options for the researcher.
The sorter is capable of up to 4 sorts simultaneously into a variety of different receptacles. Also, single cell deposition into most standard 80mm x 120 mm multi-well tray formats (e.g., 96-well plate) is routine with the ACDU system. Both sample and collection tubes can be kept at a fixed temperature (5-37oC), maximising viability.
To view fluorescence channels, go to Fluorochromes tab.
ImageStream X Mk II:
The Amnis ImageStream X Mk II (Luminex) imaging cytometer, the first such instrument installed in Ireland, is a powerful platform that combines the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy. At rates of thousands of cells per second, the instrument produces multiple high-resolution images of each individual cell directly in flow, with up to 10 fluorescent markers (405, 488, 561, & 640 nm lasers) and sensitivity exceeding conventional flow cytometers. This unique technology enables a broad range of applications that cannot be achieved with existing instrumentation within conventional flow cytometry and confocal microscopy. Beyond cellular imagery, the ImageStream is particularly suited to small particle analysis (e.g., extracellular vesicles).
In addition to the hardware, the platform is supported by powerful analysis software (IDEAS software), which combines image analysis, statistical rigour, and visual confirmation. Hundreds of parameters are calculated for every single cell, including fluorescence intensity, fluorescence localisation, cell shape and cell texture, while a wide range of statistical metrics reveal variations.
To view fluorescence channels, go to Fluorochromes tab.
Aurora:
The Aurora (Cytek Biosciences) full-spectrum analyser is the first such instrument in Ireland, for the first time enabling >25 colour cytometry analysis. With 4 lasers (405, 488, 561, & 640 nm), 3 scattering channels, and 48 fluorescence channels, the Aurora system suits simple to high-complexity applications. A paradigm shifting optical design provides unprecedented flexibility, enabling the use of a wide array of new fluorochrome combinations without reconfiguring your system for each application. The state-of-the-art optics and low-noise electronics provide excellent sensitivity and resolution, yielding outstanding performance at high sample flow rates. The end result is a system that delivers high quality data where rare and dim populations are easily resolved, regardless of assay complexity. The system in TBSI includes an Automated Sample Loader, allowing fully customisable acquisition directly from 96 well plates and 40-tube racks. SpectroFlo software offers an intuitive workflow from quality control to acquisition to data analysis.
Notable features include use of up to 30 colour panels, autofluorescence extraction, and the ability to resolve almost-identical dye combinations.
To view fluorescence channels, go to Fluorochromes tab.
LSRFortessa cytometer
The LSRFortessa cytometer (BD Biosciences) is an innovative and proven platform for multicolour analysis with its powerful fluidics systems and novel optics design, all of which work to maximize signal and enable minimum cross talk.
Features include 4 lasers (405, 488, 561, & 640 nm) and 16 fluorescence parameters, with maximal sensitivity and resolution, at up to 40,000 events per second. FACSDiva software, common across BD instrument families, controls the system, affording greater application flexibility, allowing you to easily move assays from one platform to another.
To view fluorescence channels, go to Fluorochromes tab.
FACSCanto II:
The FACSCanto II cytometer (BD Biosciences) is an easy-to-use benchtop analyser that delivers proven performance, accuracy, and high-quality results.
Features include 3 lasers (405, 488 & 640 nm) and 8 fluorescence parameters, at up to 10,000 events per second. FACSDiva software, common across BD instrument families, controls the system, affording greater application flexibility, allowing you to easily move assays from one platform to another.
To view fluorescence channels, go to Fluorochromes tab.
Accuri C6:
The Accuri C6 cytometer (BD Biosciences) is an easy-to-use, intuitive benchtop analyser with simplified acquisition and analysis software.
Features include 2 lasers (488 & 640 nm) and 4 fluorescence parameters, at up to 10,000 events per second. The Accuri C6 software is also available for off-line analysis.
To view fluorescence channels, go to Fluorochromes tab.