Flow Cytometry

The Flow Cytometry Laboratory, headed by Derek Davies, provides a high-quality flow cytometry service for the research laboratories at the Crick.

Flow cytometry is a specialised form of microscopy that provides a means of measuring cells or particles as they pass one by one through a light source, which is usually a focused laser beam. It is possible to measure physical characteristics such as cell shape and complexity by the way cells scatter light. Chemical characteristics can also be ascertained by measuring fluorescence after excitation by laser light. Any part of a cell that can be labelled with a fluorochrome can therefore be detected and quantitated by flow cytometry.

Flow cytometers may be divided into analysers and sorters. Analysis can be extremely rapid - up to 20,000 events per second - so it is an accurate way of quantifying properties of interest. In addition, it is possible to retrieve (or sort) cells identified by analytical flow cytometry for further study such as re-growth, extraction of nucleic acids or further functional analyses. Users of the facility are trained in the use of analytical cytometers while operation of cell sorters is by the laboratory staff.

Major applications within the Flow Cytometry laboratory are multicolour phenotyping (we can measure up to 17 colours on an individual cell), DNA analysis and cell cycle kinetics, cell death and apoptosis as well as functional assays such as cell killing or ion flux.

The service encourages interaction between users and service staff, and members of the team are available to advise on experimental design, interpretation and presentation of results.

The Flow Cytometry service aims to keep at the cutting edge of cytometric technology also introduce new techniques appropriate to users needs. Currently the laboratory has eight analytical cytometers and five  high-speed cell sorters along with seven permanent members of staff.

 

Projects

The Flow Cytometry Facility at the Francis Crick Institute is a dedicated scientific service offering an extensive flow cytometry analysis and sorting facility. Flow cytometry is a sophisticated form of fluorescence microscopy where cells in suspension pass one by one through a laser beam and emitted fluorescence can be captured and measured. Any part of a cell or any function of a cell that can be tagged by a fluorochrome may be measured by flow cytometry which makes it an essential technique in many biological applications.

Equipment

Analytical cytometers

The Flow Cytometry Facility has 14 analytical cytometers including two plate-reading cytometers and two with High Throughput options. These may all be user-operated and we offer a one to one training for all new users of the facility: - FACS - Calibur - Three LSRII cytometers - Four LSR Fortessa cytometers including one X-20 model - FACSVerse - FACSCanto - MACSQuant VYB - CyAn - FlowSight Imaging flow cytometer - ImageStream imaging flow cytometer

Figure 1

Figure 1. Amnis FlowSight imaging flow cytometer.

Cell sorters

These are able to retrieve up to six specifically defined populations so that cells may be recovered for re-culture, functional assays or RNA or DNA recovery. Only members of the Flow Cytometry Facility operate the sorters but experiments are scheduled and planned in close collaboration with our users. Four of the sorters are housed in Class 2 Microbiological Safety Cabinets. - Three MoFlo XDP cell sorters - Three FACS Aria cell sorters including one FACSAria Fusion - Two Influx cell sortersAvalon cell sorter

Other services

The members of the Flow Cytometry Facility are available to provide advice on the design of experiments, sourcing and supply of reagents, data analysis, presentation and interpretation as well as troubleshooting machines and experiments. We also develop and introduce new techniques and technologies that would be useful to our users. Recently we have introduced imaging flow cytometry to the Facility's capabilities with the installation of two imaging flow cytometers, the ImageStream and the FlowSight. This will enhance the range of applications that we can provide.

Downloads

LSR Guide

MACSQuant VYB Guide

Sorting Guide

Sorter Configurations

Viability Dyes Protocol

Derek Davies

Derek Davies (Lead) 

derek.davies@crick.ac.uk
+44 (0) 20 7269 3394

Graham Preece (Assistant)