Many years ago, we detected small numbers of epithelial cells (breast carcinoma cells) in sections of lymph nodes using an immunohistochemical stain for epithelial membrane antigen (EMA) (The use of antisera to epithelial membrane antigen in detecting micrometastases in histological sections. J.P. Sloane, M.G. Ormerod, S.F. Imrie and R.C. Coombes. Brit. J. Cancer, 42, 392-398, 1980). You could use the same approach in flow. Having made a single cell suspension from the node, I would label the cells with EMA (rat monoclonal antibody from Harlan Sera-Lab Ltd.) and CD45. Alternatively, you could label with CD45, permeablise and label with monoclonals to cytokeratin. I would prefer the first approach as you do not need to fix or permeabilise the cells before labelling. Michael Ormerod 34 Wray Park Road Reigate RH2 ODE Telephone: voice & FAX: (0)1737 241726 Web site: http://ourworld.compuserve.com/homepages/Michael_Ormerod
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