Flow Cytometry Facility
Manager: Dr. Alison M. Michie
Paul O’Gorman Leukaemia Research Centre
Telephone: 0141 301 7885
Email: Alison.Michie@glasgow.ac.uk
Dr. Heather G. Jørgensen
Paul O’Gorman Leukaemia Research Centre
Telephone: 0141 301 7875
Email: heather.jorgensen@glasgow.ac.uk
Operator: Jennifer Cassels
Paul O’Gorman Leukaemia Research Centre
Telephone: 0141 301 7887
Email: Jennifer.Cassels@glasgow.ac.uk
What is Flow Cytometry?
Flow cytometry is a technique that allows the characterisation and separation of particles in suspension (e.g. cells, nuclei or chromosomes) by virtue of size, granularity and fluorescence (autofluorescence or fluorescence after immunolabelling or staining). The introduction of flow cytometry technology has enabled the separation of more homogeneous, pure populations from a mixed population, thus allowing the complexity of a population to be deciphered – fluorescence activated cell scanning/sorting (FACS). Advances in cell sorting instrumentation have enabled this technology to be applied to a wider array of cell biology areas, such as haematology, immunology, microbiology, cellular signalling, and parasitology. Moreover, a number of advances in the new generation of cell sorters in sensitivity and speed of separation have allowed for a more rapid, detailed examination of molecular and cellular components.
Our Cell Sorter – BD FACSAria
The BD FACSAria cell sorter has a revolutionary design that has created a stable instrument with which to perform cell sorting. This has been achieved by introducing a number of novel features: fixed-alignment cuvette flow cell; self-contained fluidics and; improved computer software. These advances have revolutionised, while at the same time simplified, the way cell sorting can be achieved.
Our cell sorter offers a number of powerful features summarised below:
- three lasers - 407-nm violet, 488-nm argon, and 633-nm HeNe;
- multi-parameter analysis which allows up to twelve independent signals to be analysed at one time for a more detailed phenotypic analysis of cell populations;
- digital compensation to increase sensitivity of analysis;
- acquisition of up to 90,000 events/second;
- sorting of up to 25,000 events/second, with > 98% purity;
- collection of four cell samples concurrently to reduce the amount of sample required;
- sorting into a number of different collection devices, such as: microtubes, 5 ml and 15 ml tubes and 6, 12, 24, 48, 96, 384 well plates;
- isolation of single cells in tissue culture plates for cloning purposes;
- sorting of defined ratios of multiple cell types in a single-pass for experimental set-up;
- collection of rare cell events, due to the increase in sensitivity;
- temperature control of samples - the instrument offers temperature regulation (heating or cooling) of unsorted and sorted samples;
- aerosol management system for operator protection from potentially biohazardous aerosols.
The high-end configuration of our BD FACSAria places us in the unique position of owning a piece of equipment whose benefit will appreciate over the next decade as more reagents become available, thus this instrument should be considered as an investment in core research facilities within the University of Glasgow.
Our Cell Analyser – BD FACSCantoII
We also have a FACS analyser, the BD FACSCantoII, which has a similar fluorescent footprint to our FACSAria, but is tailored for analysis of specific biological properties of mixed cell populations from multiple samples.
Requests
As well as utilising these instruments to achieve our own research goals, we are dedicated to the advancement of scientific research within the University of Glasgow and beyond. Therefore if you think we may be able to assist your research please contact us and hopefully we can be of some help.
Funding
Establishment of our flow cytometry facility and service has been made possible by the following funding:
Kay Kendall Leukaemia Fund Grant
2010 – 2013
The Howat Foundation Ltd.
2010 – 2013
Useful Links
Questions and answers about cell sorting: http://cyto.mednet.ucla.edu/Protocols/question.htm
Fluorochrome excitation and emission spectra:
http://www.bdbiosciences.com/spectra/
http://pingu.salk.edu/flow/fluo.html