Lynette - Yes. We faced this problem back in 1981 when reviewers, who did not understand flow cytometry, thought that the actual distribution of viral binding was an big error around an average measurement. So we painstakingly showed the results of multiple experiments where we showed the variations in results when (1) the same sample was run multiple times, (2) the staining was done several times on aliquots of the sample. Even more painstaking was trying to explain to the reviewers that the error bars were smaller than could be shown on the graphs! They had a tough time understanding that two "overlapping curves could be statistically different. The biological variation was much, much larger than the error in measurements. Here are the old references, published in these two papers: Leary, J.F., Notter, M.F.D.: "Kinetics of Virus Adsorption to Single Cells Using Fluorescence Membrane Probes and Multiparameter Flow Cytometry" Cell Biophysics 4: 63-76 (1982). Notter, M.F.D., Leary, J.F., Balduzzi, P.C.: "Adsorption of Rous Sarcoma Virus to Genetically Susceptible and Resistant Chick Cells - A Laser Flow Cytometry Study" J Virology 41: 958-964 (1982). -- Jim James F. Leary, Ph.D. Chief, Molecular Cytometry Unit Division of Infectious Diseases 4.216 Mary Moody Northen Pavilion - Route 0435 Professor of Internal Medicine, Pathology, Microbiology & Immunology, Biophysics, Human Biological Chemistry & Genetics. and Biomedical Engineering Tel: 409-747-1930 Email: jleary@utmb.edu -----Original Message----- From: Tytherleigh, Lynette [mailto:Lynette.Tytherleigh@health.wa.gov.au] Sent: Sunday, January 18, 2004 6:28 PM To: cyto-inbox Subject: Uncertainty of Measurement Has anyone had to evaluate and document "Uncertainty of Measurement" with respect to Flow Cytometry? Any advise would be much appreciated. Regards Lynette Tytherleigh Haematology Department Fremantle Hospital Alma St Fremantle 6959 Western Australia Phone: 08 94312459 Email: Lynette.Tytherleigh@health.wa.gov.auReceived on Tue Jan 20 19:58:00 2004
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