Hello, all: We are interested in counting the number of cells expressing various cytokines in a mouse model using flow cytometry. The only published work I've seen looking at intracellular cytokines and flow was in vitro stimulation in the presence of a protein transport inhibitor. Do any of you have experience detecting intracellular cytokines from in vivo experimental sytems (such as mouse blood or spleen cells) by flow? Is it reasonable to expect to be able to get anti-cytokine antibody molecules into a cell without the cytokines leaking out? (I assume a fixation step would have to preceed the permeablization step.) I'd appreciate any information about, or directions to, a protocol which works. Thanks in advance. * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Richard K. Meister Email: Meister.1@osu.edu * * The Ohio State University Voice: (614) 292-9716 * * Dept. of Veterinary Biosciences FAX: (614) 292-6473 * * Cytometry Instrumentation Lab * * 1925 Coffey Road * * Columbus, OH 43210 U.S.A. * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id <01IHAA9602A8003K65@mc.duke.edu>; Thu, 03 Apr 1997 22:44:23 -0500 (EST) Received: from flowcyt.cyto.purdue.edu (flowcyt.cyto.purdue.edu [128.210.60.31]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id WAA07278; Thu, 03 Apr 1997 22:41:25 -0500 (EST) Received: by flowcyt.cyto.purdue.edu (940816.SGI.8.6.9/930416.SGI.AUTO) for cyto-sendout id UAA10523; Thu, 03 Apr 1997 20:35:58 -0500 Received: from mail3.uts.ohio-state.edu by flowcyt.cyto.purdue.edu via ESMTP (940816.SGI.8.6.9/930416.SGI.AUTO) for <cytometry@flowcyt.cyto.purdue.edu> id RAA07759; Thu, 03 Apr 1997 17:24:08 -0500 Received: from [128.146.83.18] (rmeister2.vet.ohio-state.edu [128.146.83.18]) by mail3.uts.ohio-state.edu (8.8.5/8.8.5) with SMTP id RAA09923 for <cytometry@flowcyt.cyto.purdue.edu>; Thu, 03 Apr 1997 17:14:14 -0500 (EST) Date: Thu, 03 Apr 1997 17:16:13 -0600 (CST) From: "Richard K. Meister" <rmeister@pop.service.ohio-state.edu> Subject: Intracellular cytokine measurement Sender: rmeister@pop.service.ohio-state.edu To: cyto-inbox Message-id: <62174.rmeister@pop.service.ohio-state.edu> Content-transfer-encoding: 7BIT X-NUPop-Charset: English
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