Dear Flow Folks, We have a user who has posed the following project: Brief Description of Experiment: I have a suspension of monodisperse microspheres, one micron in diameter. Each sphere has identical DNA strands grafted to its surface. I need to determine the absolute number of DNA's per sphere. I am currently using DNA that has a single FITC molecule attached to each DNA strand. How accurately can the flow cytometer determine the absolute number of DNA's per sphere? I should also note that my samples have approximately 10000 DNA's per sphere. How low in number can I go before the cytometer gives poor results? Aside from the usual issues of standardizing for quantitative measurements, would any of you have any advice. What would you say would be the fewest number of FITC molecules that one could detect on a standard tabletop instrument. I am a little uncomfortable with committing to this level of sensitivity, especially with FITC.. Thoughts!! thanks, Jonni -- Jonni S. Moore, Ph.D. Associate Professor of Pathology and Laboratory Medicine Director, PATH BioRESOURCE Director, Flow Cytometry and Cell Sorting Shared Resource, Abramson Cancer Center Director, Clinical Flow Cytometry, Hospital of the University of Pennsylvania University of Pennsylvania School of Medicine 220 John Morgan Bldg. Philadelphia, PA 19104-6082 Phone: 215-898-6853 FAX: 215-898-0734Received on Wed Jul 13 13:58:00 2005
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