Dear Flow-ers, We recently encountered some problems with the sort of transfected cells. When we sort GFP+ mouse embryonic stem cells (48 h after Xgen transfection with a vector expressing the gene of interest and GFP), the sorted positive cells gradually loose their fluorescence within 3 to 4 days. On the other hand the same transfected cells, selected for example in G418, retain their fluorescence much longer. Viability etc. of the sorted cells is very good. Any ideas, how this phenomenon can be explained, would be appreciated. (Was there any deleterious effect of cell sorting on long term protein expression described before?) Any proposals how to get around it would be even better. Thank you very much in advance. Jochen Dr. Jochen BARTHS IGBMC Service Commun de Cytometrie en Flux 1, rue Laurent Fries BP 10142 F-67404 ILLKIRCH CEDEX France Tel.: +33 / (0)3 / 88 65 34 27 or 28 Fax: +33 / (0)3 / 88 65 32 01 e-mail: JBarths@IGBMC.u-strasbg.fr
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