Award details

A fluorescence activated cell sorter facility for the biosciences

Reference	BB/C511172/1
Principal Investigator / Supervisor	Professor George Blair
Co-Investigators / Co-Supervisors	Professor Noel Buckley, Professor Simon Carding, Dr Jurgen Denecke, Dr MPG Harris, Dr Eric Hewitt, Professor Julian Hiscox, Professor Nigel Hooper, Professor E Ingham, Dr Kenneth McDowall, Professor Michelle Peckham, Dr Sreenivasan Ponnambalam, Professor David Rowlands, Professor Peter Stockley, Professor Anthony Turner, Professor Adrian Whitehouse, Professor Mark Wilcox
Institution	University of Leeds
Department	Inst of Molecular & Cellular Biology
Funding type	Research
Value (£)	153,581
Status	Completed
Type	Research Grant
Start date	30/04/2005
End date	29/04/2006
Duration	12 months

Abstract

The ability to identify and purify cell types forms an important strategy in many areas of current molecular and cell biology. Of the many techniques available to purify rare populations of cells, fluorescence activated cell sorting (FACS) is the most powerful, combining the precision of identification and recovery of viable cells (based on their tagging with fluorescent markers) with high throughput of cells, of the order of 10 to the power of 8 cells per hour. Cell sorters are able to sort mammalian, plant and bacterial cells. Advances in fluorescence protein technology now permit the selection of cells expressing any, or several, of a wide range of fluorescent proteins that absorb across the UV and visible spectrum, not just green fluorescent protein (GFP). The major objective of this application is to provide a state-of-the-art facility for cell separation based on flow cytometry that will have great benefit for a large group of bioscientists at Leeds, working across the entire spectrum of biological research in three Schools (Biochemistry and Microbiology, Biology and Biomedical Sciences) in the Faculty of Biology Sciences. The consequent impact on research will be great, allowing the formulation of new approaches to biological problems and an increase in productivity. Most of the projects that will benefit from this facility fall within the areas of two multi-disciplinary research groupings within the Faculty: Molecular Cell Biology and Infection and Immunity. The first area incorporates important and timely work on stem cell biology, both with respect to the identification and differentiation of neural stem cell populations and in the use of adult stem cells for tissue engineering. Stem cell research and tissue engineering are BBSRC priority areas and the neural stem cell project relates to Brain Science, a current Foresight objective. The isolation of cell lines that express defined levels of the prior protein, the causative agent of the transmissible spongiform encephalopathies, along with further investigations into its proteolytic processing also relates to Brain Science. Further projects that require cell sorting concern fundamental questions in cell biology, such as protein trafficking in mammalian endothelial cells and plant cells and motility in muscle cells. Cell sorting will enable the selection of rare transfected cells expressing fluorescent proteins such as GFP and recombinant proteins that affect the biological properties of these varied cell types. Such fundamental areas in cell biology fall within the remit of the BBSRC Biochemistry and Cell Biology Committee. The second area that will benefit from this equipment incorporates projects in Infection and Immunity. Bacterial and viral pathogenesis are important themes in the Infection and Immunity Grouping and cell sorting will be used to isolate rare populations of gamma delta T cells from pathogen-infected mice thus enabling further molecular studies to be performed on this interesting class of T cells. Several projects require sorting of cells expressing viral proteins (from HPV, HVS, HIV-1 and Rhinoviruses). Studies on viral entry require sorting of cells expressing receptors for human adenoviruses and ACE2, the recently-identified cell receptor for the SARS coronavirus. Cell sorting forms a crucial part of a novel mutatenesis and screening protocol using GFP-fusion proteins in Escherichia coli and further projects in purification of Clostridia sp. Will only be possible using a FACS sorter. We have extensive experience in analytical flow cytometry, possessing two BD FACScalibur instruments that are run as a managed core facility, offering training and advice in analytical cell biology to researchers in Leeds in Biological Sciences, Medicine, Leeds General Infirmary and groups in the University of Bradford. However, we have no capability to sort cells. Acquisition of the FACSAria would greatly expand our facility and enable us to offer a comprehensive analytical and cell sorting service to the large biological community in the Leeds/Bradford area.

Summary

unavailable

Committee	Closed Committee - Biochemistry & Cell Biology (BCB)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Research Equipment Initiative 2004 (RE4) [2004]
Funding Scheme	X – not Funded via a specific Funding Scheme

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