Introduction

Flow cytometry refers to the measurement and analysis of various phenotypical, biochemical or molecular characteristics of cells (or particles), usually labelled with fluorescent probes, as they pass one or more laser beams in single file through a fluid stream.

Analysis parameters include size, granularity and structural complexity, surface receptors, intracellular proteins and cytokines, enzymes, apoptosis, nucleic acid content, cell cycle analysis, calcium flux etc. Flow sorting extends to the enrichment/ purification of cell population subsets, which exhibit an identifiable set of scatter and fluorescent characteristics.

The Flow Cytometry Facility in Trinity College Dublin was set up in 2005 within the School of Biochemistry and Immunology. At present, it is equipped with one flow cytometer sorter and two analysers:

Beckman Coulter (Dako) 3-laser, 8 channel, MoFlo hi-speed cell sorter which can analyse and sort up to four populations at once, at speeds of >50,000 cells per second. Can sort into various receptacles (tubes, plates, slides etc.)
Beckman Coulter (Dako) 2-laser CyAn ADP analyser which can analyse cells or particles with up to 7 fluorochromes, at speeds of up to 50,000 cells per second
Becton Dickinson 1-laser FACSCalibur for routine analysis of cells with up to 3 fluorochromes.

Sorting services are provided on the MoFlo whereby the prepared samples are brought to the facility and the required populations (up to 4 populations at a time) are selected and purified. Samples for the MoFlo sorter can be stained with up to 6 fluorochromes simultaneously (see Fluorochromes). Experimental design and assistance etc. is provided.

The CyAn ADP and FACSCalibur cytometer are available to trained operators. Training and/or experimental and technical assistance is provided. Data analysis may be performed on-site with Flowjo, Summit, or CellQuest software, or off-site with various analysis packages.

Sorting services, and use of the cytometers, is charged at an hourly rate, with prices varying for TCD, academic and industrial researchers.