Flow cytometry

Overview

Flow Cytometry is a technology that simultaneously measures multiple characteristics of a single cell at a rapid rate while in a moving fluid stream. It can tell us about physical characteristics such as cells' relative size, granularity or internal complexity and any cell component or function that can be detected by a fluorescent compound.

The main applications of flow cytometry include immunophenotyping, intracellular protein staining, cell viability, cell tracking and proliferation, DNA cell cycle and degradation, cell apoptosis, enzyme activity, phosphorylation, intracellular calcium influx and cell sorting.

Having identified a specific subpopulation using an analytical cytometer, flow cell sorting allows physical separation of the particle or cells of interest with high purity from a heterogeneous cell population for further study.

Flow Cytometry at the Kennedy

The Kennedy flow facility is located on the first floor in room 20.22 (the analyser room) and room 20.26 (the sorting room).

The facility is equipped with state-of-the-art instruments that provide an exciting and innovative research and training environment for researchers to carry out immune monitoring on tissue and blood samples.

The facility provides investigators with instruments and support for cell sorting and acquisition. The facility is managed by Jonathan Webber and directed by the academic lead for the facility, Prof Michael Dustin. The Kennedy Trust has funded this facility.

Cell Analysis

The facility holds three bench top FACS analysers:

• BD SORP LSRFortessa X20 with HTS (high throughput sampler - plate reader)
• BD LSR II
• Cytek Aurora (5L 64-detector spectral cytometer with HTS)

Cell Sorting

The facility holds two FACSAria III cell sorters:

• KIR four lasers (belongs to the Kennedy Institute of Rheumatology, in a cat 1 BSC)
• ORC three lasers (belongs to the Department of Oncology at the old road campus

Other

Meso Quickplex SQ120 - a plate reader for the MSD mutliplexed imunnoassay kits. Up to 10 simultaneous analyte measurements.

Services

The facility staff are able to provide advice and support with the following:

• Training on how to operate cell analysers
• Experimental design
• Panel design for multicolour FACS analysis
• Compensation and sample acquisition
• Sample preparations for cell sorting and acquisition
• Cell sorting
• Data analysis