In the Department of Surgery
University of Newcastle.
About Us
Equipment
Specific Research Interests
General Techniques Available
Description and Users of the Facility
Flow Cytometry Courses
Useful Links
About Us
A Brief History
In 1983, it was proposed by the Transplantation Immunology Research Group in the Department of Surgery that the application of flow cytometric technology to problems in the area of Renal Transplantation had tremendous potential research value. After consultation with colleagues, it became clear that the research potential of such technology was not restricted to Transplantation and considerable support was given by workers within the University Departments of Pathology, Immunology and Haematology. A local appeal was therefore launched by Mr. George Proud and Dr. BK Shenton and in 9 months was successful in raising the required money to purchase a FACS 420 Flow Cytometer. Once acquired the facility was offered to other University and Hospital Departments. Consequent upon such humble beginnings the Flow Cytometer has achieved crucial importance in many research projects funded by the Wellcome Trust, the Medical Research Council, the British Heart Foundation and many local charities including the Northern Counties Kidney Research Fund, the North East Cancer Research Campaign, the Womans Cancer Detection Society and District and Regional Hospital grant awarding bodies. To help run the Flow Cytometric facility Dr. Alice Givan was employed for 2 years before she left for her present position in the States. From initial research studies data from flow cytometry has significantly altered the management of renal and cardiothoracic transplant patients. The two main areas which have received international recognition have been those of immunosuppressive monitoring and development of the Flow Cytometric Crossmatch test. In 1983 with the availability of Bench Top Flow Cytometers Dr. Brian Shenton applied to the Great North Run Charity through Mr. Ross Taylor for money to buy a FACScan Flow cytometer. Within three months the money was raised and the Flow Cytometer was presented to the Department by Brendan Foster on behalf of the Great North Run Charity. Again use of the facility outgrew the availability and in 1995 Dr Brian Shenton applied to the Northern Counties Kidney Research Fund for another Bench Top Flow Cytometer - the FACSort. This was presented six months later. Since 1993, the use of the FACS 420 cell sorter has diminished and the equipment was removed in 1998, prior to the arrival of the Coulter Elite Cell Sorter.
The flow cytometry facility has been run by Dr Shenton since 1983, and by arrangement both advice and help are provided for research and technical staff in the University, the Hospitals and the Northern Region. Assistance for this is provided by Alison Bell.
How to find us:
The Flow Cytometers are situated in the Department of Surgery (Room M308, level 3 of the William Leech building, The Medical School, Framlington Place, University of Newcastle, NE2 4HH.
For further information contact:
Dr B.K. Shenton or Alison Bell
Telephone (0191) 222 7079 or (0191) 2228514
Fax (0191) 222 8514 or (0191) 222 8441
Email B.K.Shenton@ncl.ac.uk
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The flow cytometry laboratory houses :
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Breast Cancer
Gastrointestinal Cancer
Transplantation
Vascular Immunology
Research Group
Dr B.K. Shenton (Consultant Clinical Scientist)
Ms Fiona Girdler (PhD student commencing 1998)
Mr Alex Navarro (BMedSci Student commencing 1998)
Clinical Group
Mr D.A. Browell (Surgical Director, QE Hospital, Gateshead)
Mr W.J. Cunliffe (Consultant Surgeon, QE Hospital, Gateshead)
Mr M.J. Higgs (Consultant Surgeon, QE Hospital, Gateshead)
Mr TWJ Lennard (Head of Dept of Surgery, Medical School)
Mr R. Bliss (Consultant Surgeon,QE Hospital, Gateshead)
Mr A. Griffiths (Consultant Surgeon, RVI)
Mr C. Griffiths (Consultant Surgeon, RVI)
Dr L.G. Lunt (Consultant Radiologist, QE Hospital, Gateshead)
Dr L.A. Webb (Consultant Radiologist, QE Hospital, Gateshead)
Dr J.R. Young (Medical Director, Breast Screening Unit, QE)
Research Interests
Use of Fine Needle Aspirates for diagnostic purposes by flow cytometry.
Use of flow cytometry on solid primary tumour to examine prognostic markers.
Isolation of individual tumour phenotypes.
The effect of endocrine therapy on growth receptors and adhesion molecules.
Isolation of tumour cells by sorting, for molecular biological analysis.
Examination of cellular structural proteins.
Cell cycle, DNA and Apoptotic markers.
Selected Publications.
Brotherick, I.,et al. Flow cytometric method for the measurement of epidermal growth factor receptor and a comparison with the radio-ligand binding assay. Cytometry 16, 262-269, 1994.
Brotherick, I., et al. Association between c-erbB-2 proto oncogene expression, grade and ploidy in breast tumours. Cancer Immunol. Immunother., 41, 137-145, 1995.
Brotherick, I.,et al. Use of the biotinylated antibody DAKO-ER 1D5 to measure oestrogen receptor on cytokeratin positive cells obtained from primary breast cancer cells. Cytometry, 70,74-80, 1995.
Brotherick, I., et al. p53 expression measured by flow cytometry. A comparison of three monoclonal antibodies and the relationship with grade and DNA ploidy in breast cancer. Cancer Immunol. Immunother., 41, 146-150, 1995.
Brotherick, I., et al. A flow cytometric study of c-erbB-3 expression in breast cancer. Cancer Immunol. Immunother., 41,280-286, 1995.
Brotherick, I., et al. Are fine needle aspirates representative of the underlying solid tumour? Br J Cancer, 72, 732-737, 1995.
Brotherick, I.,et al. Examination of multidrug resistance in cell lines and primary breast tumours by flow cytometry. Eur. J. Cancer, 32A, 13, 2334-2341, 1996.
Brotherick, I., et al. The Effect of Three Week Tamoxifen Treatment on
Oestrogen Receptor Levels in Primary Breast Tumours: A Flow Cytometric Study. Br. J. Cancer, 77, 10, 1657-1660, 1998.
Brotherick, I., et al. Cytokeratin expression in Breast Cancer: Phenotypic Changes associated with disease progression. Cytometry, 32, 301-308, 1998.
Supported by The Women's Cancer Detection Society (WCDS), Gateshead.
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The applications of Flow Cytometry to Gastrointestinal Cancer research has developed as a collaboration between a variety of groups. Six main MD's are using the flow cytometry facility for colorectal projects.
Researchers
Mr P. Bhaskar (The role of growth factors and their receptors in acromegalic polyps)
Mr J.M. Hanson (Mucin expression)
Mr D. Karat (Hormone receptors in gastric cancer)
Mr K. Seymour (The importance of adhesion molecules in gastro-intestinal cancer)
Mr D.S. Watson (Proliferation and Apoptosis markers)
in the Laboratory:-
Dr B.K. Shenton (Consultant Clinical Scientist)
Clinical Collaborators
Mr R. Charnley (Consultant Surgeon at Freeman Road Hospital)
Mr S.M. Griffin (Consultant Surgeon at the Royal Victoria Infirmary)
Mr S. Raimes (Consultant Surgeon at Carlisle General Hospital)
Mr J. Varma (Senior Lecturer in General Surgery, Royal Victoria Infirmary)
Mr S. Kelly (Senior Lecturer in General Surgery, North Tyneside)
Research Interests
Cyclin expression and proliferation.
Adhesion molecules.
Oestrogen receptor.
Progesterone Receptors.
Insulin like growth factors.
Mucins 1,2 and 5ac.
Cell cycle, DNA and Apoptotic markers.
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Transplantation
Transplantation has been the major use for flow cytometry since the first flow cytometer was acquired in the Department of Surgery and its application and development is lead by Dr B.K. Shenton. The group has strong clinical links with the RVI and Freeman Hospitals as well as the Blood Transfusion Service. Considerable research has centered around Kidney, Heart, Lung and Liver Transplantation.
Research Group
Dr B.K. Shenton (Consultant Clinical Scientist)
Mrs A.E. Bell (MPhil, Research Technician)
Mr G. Cavanagh (Senior Clinical Scientist, Blood Transfusion Service, Newcastle)
Mr V. Carter (Senior Technician, Blood Transfusion Service, Newcastle)
Clinical Group
Mr D. Manas (Consultant Surgeon, Freeman Hospital)
Mr D. Talbot (Consultant Surgeon, Freeman)
Mr J. Dark (Consultant Surgeon, Freeman Hospital)
Research Interests
FACS Crossmatch against Donor Cells.
Crossmatch against Cell lines and specificity of donor antibodies before and after transplantation.
Monitoring of the efficacy of ATG and ALG therapy
Detection of antibodies against ATG and ALG in transplant and patients with Aplastic Anaemia
Phenotyping for activation markers
Quality control of flow cytometry in Tissue Typing Departments throughout the United Kingdom
Importance of adhesion molecules in transplantation and the role of monoclonal antibodies as therapy for rejection.
Talbot, D., Givan, A.L., Shenton, B.K., Proud, G. & Taylor, R.M.R. Transplant Proc 19, 4315-4316 (1987).
Talbot, D., Shenton, B.K., Givan, A.L., Proud, G. & Taylor, R.M.R. J Immunol Methods 99, 137-140 (1987).
Talbot, D., Stratton, A., Shenton, B.K., Proud, G. & Taylor, R.M.R. Transplant Proc 20, 382-383 (1988).
Talbot, D., Givan, A.L., Shenton, B.K., Stratton, A., Proud, G. & Taylor, R.M.R. Transplant. 47, 552-555 (1989).
Talbot, D., Givan, A.L., Shenton, B.K., Proud, G. & Taylor, R.M.R. Transplant 47, 552-555 (1989).
Talbot, D., Givan, A.L., Shenton, B.K., Proud, G. & Taylor, R.M.R. Transplant Proc 20, 84-85 (1989).
Shenton, B.K., Parrott, N.R., Givan, A.L., et al. Transplantation and Clinical Immunology 21, 229-229 (1989).
Shenton, B.K., Givan, A.L., Wilson, R.G., et al. Transplant Proc 20, 762-763 (1989).
Givan, A.L., Shenton, B.K., White, M.D., Proud, G. & Taylor, R.M.R. Clin.Immunol.Immunopathol. 54, 419-430 (1990).
Shenton, B.K. Micro 90 203-206 (1990).
Shenton, B.K., Glenville, B.E., Mitcheson, A.E., et al. Transplant Proc 23, 1153-1154 (1991).
Talbot, D., Cavanagh, G., Coates, E., et al. Transplant. 53, 925-928 (1992).
Talbot, D., Shenton, B.K., Givan, A.L., Cavanagh, G., Proud, G. & Taylor, R.M.R. Transplant Int 5, S604-S605(1992).
Clark, K.R., Forsythe, J.L.R., Shenton, B.K., Proud, G. & Taylor, R.M.R. Transplantation and Clinical Immunology 24, 217-220 (1992).
Kutuckculer,N, Shenton, B.K. & White, M.D. Turkish Journal of Medical science 18, 219-226 (1993).
Clark, K.R., Forsythe, J.L.R., Shenton, B.K., Lennard, T.W., Proud, G. & Taylor, R.M.R. Transpl.Int. 6, 18-21 (1993).
Al Hussein, K., Shenton, B.K., Bell, A., et al. Transplant Proc 25, 259-261 (1993).
Kutukculer, N., Mir, S., Shenton, B.K., Mitcheson, A. & Kutukculer, A. Diyaliz,Transplantasyon ve Yanuk/Dialysis 7, 9-13 (1993).
Shenton, B.K., White, M.D., Bell, A.E., et al. Transp Proceed 26, 3177-3180 (1994).
Shenton, B.K. Transplant Immunology 2, 135-137 (1994).
Shenton, B.K., Bal, W., Bell, A.M., Bookless, B., Corris, P. & Dark, J.H. Transplantation International 7 (suppl 1), S359-S362(1994).
Al Hussein, K., Shenton, B.K., Bell, A., et al. Transplant International 7, 182-189 (1994).
Shenton, B.K., White, M.D., Bell, A.E., et al. Transplant Proc 26, 3177-3180 (1994).
Shenton, B.K., Bal, W., Bookless, B., et al. Transpl Proceed 27, 1295-1297 (1995).
Talbot, D., Bell, A.E., Shenton, B.K., et al. Transplant 59, 737-740 (1995).
Al Hussein, K., Talbot, D., Proud, G., Taylor, R.M.R. & Shenton, B.K. Transplantation Intrnational 8, 214-220 (1995).
Talbot, D., Shenton, B.K., Bell, A., et al. Transplant Proceed 28, (6):3212-3213 (1996).
Harmer, A., Garner, S., Bell, A.E., et al. Transplant 61, 1108-1111 (1996).
Shenton, B.K. Proceedings of the Royal Microscopical Society 31, 132(1996).
Shenton, B.K., Bal, W., Bell, A.E., et al. Transp Proceed 27, 1295-1297 (1997).
Shenton, B.K., Bell, A., Boyce, M., et al. Transplant Proc 29, 1454-1455 (1997).
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Dr B.K. Shenton
Mr B. Bannergee
Mr B. Akolomalafe
Clinical Group
Mr D.Lambert (Consultant Surgeon, Freeman Hospital)
Mr T. Lees (Consultant Surgeon, Freeman Hospital)
Dr P. Kestevan (Consultant Haematologist, Freeman Hospital)
Research Interests
The response of polymorphonuclear leucocytes to exercise in patients with vascular disease.
The relationship between polymorphonuclear leucocyte and platelet activation in patients with varicose veins.
The value of platelet function tests in haematological patients.
The effect of cardiac bypass surgery on polymorphonuclear leucocyte and platelet function.
Standardization of platelet and polymorphonuclear function tests.
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Immunophenotyping - Dr B.K. Shenton, Mrs A.E. Bell
Antigen Quantification - Dr B.K. Shenton, Mrs A.E. Bell
Quality Control - Dr B.K. Shenton, Mrs A.E. Bell
Analysis of DNA - Dr I. Brotherick & Dr B.K. Shenton
Assessment of Apoptosis- Dr I. Brotherick
Flow Cytometric Crossmatching - Dr B.K. Shenton, Mrs A.E. Bell
ATG monitoring - Dr B.K. Shenton, Mrs A.E. Bell
Measurement of internal cellular proteins -Dr B.K. Shenton
Bench Top Flow Cytometric Instrument Set Up and Calibration Dr B.K. Shenton.
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What is the facility?
The facility in the Department of Surgery is to allow any users to discuss the applications of flow cytometry to their research work. This involves the use of all facilities except the Elite Cell Sorter available in the Transplant laboratory of the Department of Surgery. All interested workers please contact Brian Shenton in the Transplant Laboratory - phone or fax 0191-2228441 or e-mail b.k.shenton@ncl.ac.uk
Current Departmental Users in addition to those listed above.
1. Transplant Immunology Group -
Dr. J.A. Kirby, Dr S. Ali, Mr S. Petit, Miss S. Fritchley, Mr E.'O Flaherty.
2. Prostate Cancer Group
Dr C.N. Robson and Dr A. Collins
3. Head and Neck Cancer
Mr J.P. Jeannon.
Extra Departmental Users.
Dr J. Noble - Phenotype expression on Neutrophils with age.
Mrs C. Kourvelou and C. Gibbs - Phospholipid asymmetry in dialysis patients
Mr P. Senior - CD69 and CD11b expression on neutrophils.
Mr J. Harris Apoptosis Department of Dentistry, University of Newcastle.
Dr E. Routledge - Virology
Dr G. Tomms - Virology
Prof Self - Clinical Biochemistry
Novocastra
Cost
In order to recover the costs of the maintenance of the FACScan, FACSort and the Cell Washer it is necessary that we make a charge to users for the facility. This is currently £30-00 per hour or £1750 for unlimited use over a one year period. Users are encouraged to discuss all new protocols and costings with Brian Shenton, if possible before grant applications are made.
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There are two types of Course that are provided for new or established
users of the Facility.
1. A Biennial Benchtop flow cytometry course , which is extended over two days and includes demonstrations, wet workshops, presentations and discussions. This was last held at the end of July 1998.
2. In house flow cytometry courses are organized by Brian Shenton every 18 months.
For details of courses available both locally and nationally please contact Brian Shenton.
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UWCM Haematology Cytonet UK
http://www.uwcm.ac.uk/uwcm/hg/hoy/index.html
ICRF http://www.icnet.uk/axp/facs/davies/links.html
ISAC Home Page http://nucleus.immunol.washington.edu/ISAC.html
Wiley Cytometry web site http://journals.wiley.com/cytometry/
Flow Cytometry on the Web http://pingu.salk.edu/fcm/sitelink.html
Beckman Coulter http://134.217.3.35/
ACS Web Page http://www.enterprise.net/appcysys/
Becton Dickinson http://www.bdfacs.com
Dako Ltd http://www.dakoltd.co.uk
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