Dear Flowers, I am having some problems in detecting SP cells isolated from muscle. We have been successful in identifying these cells in other tissues, but for some reason in our hands cells from muscle do not stain well with Hoechst (only 10-30% of cells are stained), and I can't see that nice tail. Instrument is a MoFlo, with Enterprise laser, which we use at 25 mW. Filters are 450/20 for Hoechst blue, 680LP for H red, and 610SP is used to split emissions. The protocol we follow is the one described by Gooddell (to the letter!). Any hint will be greatly appreciated. Thanks Giovanna ------------------------------------------------ Giovanna Borsellino, M.D., Ph.D. Neuroimmunology IRCCS Santa Lucia Via Ardeatina 354 00179 Roma Italy Tel. ++39.06.51501521 (office) ++39.06.51501552 (lab) Fax ++39.06.51501553 e-mail: g.borsellino@hsantalucia.it gborsel@tin.it
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