Platforms: Flow Cytometry
Introduction
- The APC Microbiome Ireland Flow Cytometry Platform is located in Room 5.19, 5thFloor, Biosciences building, UCC.
- The Platform provides access to state of the art Flow cytometry instrumentation, expertise and training allowing the analysis of single cells or particles (bacteria, yeast, beads ect).
- Flow cytometry can be used for the analysis of apoptosis, cell cycle, cell signalling, DNA content, phagocytosis, cytokine production.
- Measurements are extremely rapid – up to 20,000 particles per second with up to 12 fluorescent parameters plus 2 physical parameters analysed on each cell.
- The core facility also provides access to flow cytometry analysis software (FlowJo, Diva, FCS Express) for accurate quantification of all parameters of interest.
Fluorescent Activated Cell Sorting (FACS) allows the physical sorting of cells of interest for further downstream analysis (functional assays, cloning, culture, transcriptomics, proteomics or microscopic analysis).
Instrumentation
FACS Celesta (BD):
3 laser analyser (405, 488, 640 nm), capable of measuring up to 12 fluorescent parameters.
FACS Aria Fusion (BD):
Multicolour high-speed cell sorter, equipped with 4 lasers (405 488, 561, 640 nm) and 12 detectors to measure up to 10 fluorescent parameters. The instrument is capable of the simultaneous sorting of four populations into tubes or plates. The sorter is contained in a class II A2 biosafety cabinet allowing sterile sorting and operator safety.
10X Chromium Controller
Single cell encapsulation and barcoding through the 10X chromium controller, a state-of-the-art automated system capable of encapsulating tens of thousands of cells in a matter of minutes. In addition to transcriptional readouts, the 10x system can be configured to simultaneously provide information on protein expression, chromatin configuration, and/or immune clonotype.
Please contact Platform Manager: Tadhg Crowley (tadhgcrowley@ucc.ie ) to start planning your single-cell seq experiment.
Services
- Training for novice and intermediate users on the use of flow cytometry instruments and post-acquisition data analysis.
- Training and guidance on all aspects of experimental design for flow cytometry experiments.
- Assisted running of the cell sorter is offered following consultation with the platform manager tadhgcrowley@ucc.ie. Single cell index sorting, cell type enrichment and high purity bulk sorts are available.
Contacts
Platform Manager: Tadhg Crowley Msc (tadhgcrowley@ucc.ie)
Director: Ken Nally Ph.D.
Log In here (hyperlink to https://openiris.io/Landing/Provider/2028) register to APC Flow Cytometry Platform booking and invoicing system.
