I routinely stain cells in 96 U or V-bottomed well plates (>50,000 cells + 35microL of the staining dilution). Plates are washed (2X) using 150uL of wash buffer, spun at 1,800 rpm for 2 min at 4C, then flick the supernatants directly into the sink. Cell pellets are disrupted by vortexing the plates before resuspending the cells with buffer or satining preparations. Maxime "Reed, Doug S Dr USAMRIID" <Doug.Reed@DET.AMEDD.ARMY.MIL> le 10/12/2001 21:53:17 Pour : Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> cc : (ccc : MailingList/general/Biocytex) Objet : multiwell autosampler - washing We've just gotten in our multiwell autosampler and I'm looking for some advice from those who have been using them for a while. I don't (at this point) need any help with the system itself, but what I am interested in is how people are washing the cells in the plates. Do you do it manually or are you washing with an ELISA washer. And how do you get rid of the supernatant between washes? Thanks! Doug Douglas S. Reed, Ph.D. Principal Investigator Respiratory and Mucosal Immunity Department of Aerobiology & Product Evaluation Division of Toxinology & Aerobiology U.S. Army Medical Research Institute of Infectious Diseases 1425 Porter St. Ft. Detrick Frederick, MD 21702-5011 301-619-6728 301-619-6911 (Fax) doug.reed@det.amedd.army.mil
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