Flow Cytometry Facility
Introduction
Flow cytometry is a complex technology, analysing multiple parameters on single cells at incredibly high speeds. The technology underpins biomedical science, yielding phenotypical and functional detail in cells at rates beyond 10,000 per second. It involves labelling cells with fluorescent probes, and requires high-value, sensitive, precision-based cellular analysers, along with post-acquisition analysis. This analysis may be reasonably straightforward or elaborate; for example, high-dimensional cytometry can provide up to a billion data points per single cell.
The Flow Cytometry Facility within the School of Biochemistry and Immunology was set up in 2005 and is located in the Trinity Biomedical Sciences Institute (TBSI). This best-in-class facility is the best equipped flow core in Ireland with one state-of-the-art cell sorter (FACSAria Fusion), an imaging cytometer (ImageStream X Mark II), a full-spectrum analyser (Aurora), four flow cytometers (LSRFortessa, FACSCanto II, Accuri C6 & Attune NxT), and an in vivo fluorescence & luminescence imager (PhotonImager; housed in the Comparative Medicine Unit).
- FACSAria Fusion (BD): a 4-laser, 18 parameter high-speed cell sorter. Capable of physically separating cells at rates up to 20,000 per second into ≤ 4 populations at once. Cells may be sorted into a number of different tubes and plates.
- ImageStream X Mk II (Amnis/ Luminex): a 4-laser multispectral, imaging flow cytometer designed for the acquisition of up to 12 channels of cellular imagery.
- Aurora (Cytek Biosciences): a full-spectrum analyser with 4 lasers, 3 scattering channels, and 48 fluorescence channels, enabling high-speed, highly sensitive single cell analysis.
- LSRFortessa (BD): a 4-laser cytometer, capable of analysing up to 16 fluorescence parameters at once, at acquisition rates of up to 40,000 events per second.
- FACSCanto II (BD): a 3-laser cytometer, capable of analysing up to 8 fluorescence parameters at once, at acquisition rates of up to 10,000 events per second.
- Accuri C6 (BD): a 2-laser cytometer, capable of analysing up to 4 fluorescence parameters at once, at acquisition rates of up to 10,000 events per second.
- Attune NxT (Thermo Fisher): a 3-laser cytometer, capable of analysing up to 11 fluorescence parameters at once, with sample-throughput rates of up to 10 times faster than traditional systems.
With users from over 100 research groups, both academic and industry, this is a very busy facility with extensive experience in many areas, particularly immunophenotyping, intracellular cytokine analysis, cell cycle and apoptosis studies. With a dedicated manager (Dr Barry Moran) and Research Assistant, training, supervision, and assistance is provided in all aspects; from experimental design through instrument set up, data acquisition and analysis. Although post-acquisition analysis is usually performed off-site, there is an Analysis PC available within the facility with FlowJo, SpectroFlo, IDEAS and Accuri C6 software.
All researchers must be trained on the cytometers before access (irrespective of experience). Training is run every 2-3 months.
Sorting services are provided whereby the prepared samples (and controls) are brought to the facility and the populations are selected and purified. Training is not required, and experimental design and assistance are provided.
Sorting services and cytometer usage is charged at an hourly rate, with prices varying for TBSI, academic, and industrial researchers.
Any formal presentations or publications resulting from work performed in the facility should be acknowledged. Example: “Flow cytometry and cell sorting was performed at the Science Foundation Ireland funded Flow Cytometry Facility at Trinity Biomedical Sciences Institute.”
If substantial input into experimental design, cell sorting or data analysis has been provided then co-authorship should be considered in subsequent publications.