Hello list, I have a mouse construct which expresses B-gal within a specific subset of neurons - I now want to sort a pure population of these cells via flow. So far I've had limited success using FDG and FDG derivatives - with maintenance of viability, and high background appearing to be the major problems. I'm currently attempting to optimize aspects of the procedures - e.g., loading time, tonicity etc. In the meantime, has anyone had experience with sorting neurons or loading FDG into B-gal neurons? Are there any specific problems you can draw my attention to? In addition, we are contemplating creating another mouse for a similar neuron-specific-sorting project, but this time containing an alternative reporter system. Does anyone have any opinion on what reporter might be optimal for such a procedure - GFP? luciferase? - or should I stick with B-gal? All opinions will be gratefully received. Tanya ============================================ Tanya Barrett National Institute on Aging, Gerontology Research Center, 5600 Nathan Shock Drive, Baltimore, Maryland 21224 e-mail: barrettT@grc.nia.nih.gov Tel: (410) 558 8372 ============================================
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