Platforms: Flow Cytometry

Introduction

The APC Microbiome Ireland Flow Cytometry Platform is located in Room 5.19, 5^thFloor, Biosciences building at UCC.
The Platform provides centralised state of the art Flow cytometry instrumentation, expertise and training allowing users to perform measurements of different physical and chemical characteristics on single cells or particles (bacteria, yeast, and beads) as they pass one by one through a focused laser beam.
Flow cytometry can be used for many different applications (apoptosis, cell cycle, cell signalling, FRET, DNA content, phagocytosis, cytokine production etc.).
Measurements are extremely rapid – up to 20,000 particles per second and significant multiplexing is possible (up to 12 fluorescent parameters plus 2 scatter parameters analysed on each cell).
Statistical analysis of data is done using Flow cytometry analysis software (FlowJo, Diva, FCS Express) for accurate quantification of all parameters of interest.
Fluorescent Activated Cell Sorting (FACS) allows isolation of particles of interest for further downstream analysis (functional assays, cloning, culture, transcriptomics, proteomics or microscopic analysis).

FACS Calibur (BD): 2 laser analyser (488, 635 nm), capable to measure up to four fluorescent parameters.
FACS Accuri C6 (BD): 2 laser analyser (488, 640 nm), capable to measure up to four fluorescent parameters.
FACS Celesta (BD): 3 laser analyser (405, 488, 640 nm), capable to measure up to 12 fluorescent parameters.

FACS Aria Fusion (BD):Multicolour high-speed cell sorter, equipped with 3 lasers (488, 561, 640 nm) and 12 detectors to measure up to 10 fluorescent parameters. The instrument is capable of sorting cells from up to four populations at once into a number of different tubes/ plates/ slides. The sorter is contained in a class II A2 biosafety cabinet allowing sterile sorting.

InCell Analyser (GE Healthcare): IN Cell Analyser 1000 is a modular, cellular and subcellular imaging system for fast, automated imaging and analysis in fixed and live cells.

Training for new 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.
Complex multi-colour analysis of cell populations based on light scatter, fluorescent content and intensity. Assisted acquisition in user operated FACS analysers is offered upon scheduling appointment.
Full service sorting of individual cell populations from heterogeneous samples. Single cell index sorting, particle enrichment and high purity bulk sorts also available.
On line booking system available through iLabs