Return-path: Received: from mumnode3.mtnl.net.in ([202.159.228.4]) by pop.mtnl.net.in (iPlanet Messaging Server 5.2 Patch 2 (built Jul 14 2004)) with ESMTP id <0IZ200KUVJLN95@pop.mtnl.net.in> for modi@mtnl.net.in; Thu, 11 May 2006 02:57:23 +0530 (IST) Received: from mx1.mtnl.net.in ([202.159.228.19]) by mx.mtnl.net.in (iPlanet Messaging Server 5.2 Patch 2 (built Jul 14 2004)) with ESMTP id <0IZ200MBTJLN2G@mx.mtnl.net.in> for modi@mtnl.net.in (ORCPT modi@mtnl.net.in); Thu, 11 May 2006 02:57:23 +0530 (IST) Received: from flowcyt.cyto.purdue.edu (128.210.61.51) by mtnl-av (202.159.228.19); Thu, 11 May 2006 02:48:42 +0530 Received: (from root@localhost) by flowcyt.cyto.purdue.edu (8.12.10/8.12.10) id k4AFnAVZ081086 for cyto-sendout; Wed, 10 May 2006 11:49:10 -0400 (EDT envelope-from owner-cytometry) Received: from flowcyt.cyto.purdue.edu (localhost.cyto.purdue.edu [127.0.0.1]) by flowcyt.cyto.purdue.edu (8.12.10/8.12.10) with ESMTP id k49JVpQ4060401 for ; Tue, 09 May 2006 14:31:51 -0500 Received: (from mailnull@localhost) by flowcyt.cyto.purdue.edu (8.12.10/8.12.10/Submit) id k49JVpEL060400 for cytolist; Tue, 09 May 2006 15:31:51 -0400 (EDT envelope-from mailnull) Received: from mtaout5.barak.net.il (mtaout5.barak.net.il [212.150.49.175]) by flowcyt.cyto.purdue.edu (8.12.10/8.12.10) with ESMTP id k49JVm7s060382 for ; Tue, 09 May 2006 15:31:48 -0400 (EDT envelope-from utk1@013.net) Received: from utk ([128.139.104.106]) by mtaout5.barak.net.il (Sun Java System Messaging Server 6.1 HotFix 0.02 (built Aug 25 2004)) with ESMTPA id <0IZ0007NAJTB5JD0@mtaout5.barak.net.il> for cytometry@flowcyt.cyto.purdue.edu; Tue, 09 May 2006 22:36:47 +0300 (IDT) Content-return: prohibited Date: Tue, 09 May 2006 22:28:10 +0200 From: Uriel TK Subject: Re: Magnetic cell sorting To: cyto-inbox Reply-to: Uriel TK Message-id: <00cb01c673a7$174bbcd0$0300000a@utk> MIME-version: 1.0 X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 X-Mailer: Microsoft Outlook Express 6.00.2900.2869 Content-type: multipart/mixed; boundary="Boundary_(ID_EgBpgB4RkcTUgZul/Gl3Zg)" X-Priority: 3 X-MSMail-priority: Normal X-eSCM-MailFrom: X-Spam-Score: -4.8 () BAYES_00,HTML_MESSAGE X-Scanned-By: MIMEDefang 2.39 X-PMFLAGS: 570950016 0 1 60406.cnm X-Antivirus: AVG for E-mail 7.1.392 [268.5.5/333] References: <20060506075054.50562.qmail@web36204.mail.mud.yahoo.com> <957311f5693dc99d0fce116a25f48d58@umdnj.edu> Original-recipient: rfc822;modi@mtnl.net.in This is a multi-part message in MIME format. --Boundary_(ID_EgBpgB4RkcTUgZul/Gl3Zg) Content-type: multipart/alternative; boundary="Boundary_(ID_H5SqE1sdOHoKMM4Hg5z76Q)" --Boundary_(ID_H5SqE1sdOHoKMM4Hg5z76Q) Content-type: text/plain; charset=iso-8859-1 Content-transfer-encoding: quoted-printable Mag-Flowers: We used to use miltenyi's system for years for CD14 separation from = peripheral blood. one year ago we did a triple comparison vs BD's new = imag system and stemcells' system. We found BD to be superior, giving = purity as good as miltenyi's, better yields, and lower cell death after = recovery. Maybe you can ask them for a sample to try out, we are very = happy with BD since we moved, and even got a deal quite a bit cheaper = than miltenyi's. good luck! Uriel. "The bitterest tears shed over graves are for words left unsaid and = deeds left undone." Harriet Beecher Stowe ----- Original Message -----=20 From: Beverly Barton=20 To: Cytometry Mailing List=20 Sent: Tuesday, May 09, 2006 1:59 PM Subject: Re: Magnetic cell sorting Hah, you are lucky, for mouse the yield is more like 10% and that's = what Miltenyi tells you to expect with their system! Sad but true..... Beverly Beverly E. Barton, Ph.D. Assistant Professor Department of Surgery/Division of Urology UMDNJ-NJMS MSB G519 185 S. Orange Avenue Newark, New Jersey 07103 Telephone 973-972-0662 E-mail bartonbe@umdnj.edu Telefacsimile 973-972-3892 On May 6, 2006, at 3:50 AM, RAJA ALAMELU wrote: Dear Flow list members, =20 My question is not directly related to flow cytometry, but now and = then I have seen questions in other areas also pop up. =20 We are attempting to do CD4 and CD8 cells positive isolation from = human PBMC, followed by Detach-a bead step, using Dynal magnetic beads. = We have followed the manufacturers instructions as it, is as well as = varied each of the conditions for optimization. =20 While the purity and viability after separation seem to be good, the = final cell yield never crosses ~30% of the total available CD4/CD8 in = PBMC. Also the initial input number, magnet bound cells and unbound = cells do not tally. =20 Is 25-30% the commonly expected yield using this technique? If = anybody has observed better yield, purity and viability, can you please = share the procedure? =20 Thanking you in advance, Alamelu Raja Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great = rates starting at 1=A2/min.= --Boundary_(ID_H5SqE1sdOHoKMM4Hg5z76Q) Content-type: text/html; charset=iso-8859-1 Content-transfer-encoding: quoted-printable
Mag-Flowers:
We used to use miltenyi's system for = years for CD14=20 separation from peripheral blood. one year ago we did a triple=20 comparison vs BD's new imag system and stemcells' system. We found = BD to be=20 superior, giving purity as good as miltenyi's, better yields, and lower = cell=20 death after recovery. Maybe you can ask them for a sample to try out, we = are=20 very happy with BD since we moved, and even got a deal quite a bit = cheaper than=20 miltenyi's.
 
good luck!
 
Uriel.
 
"The bitterest tears shed over graves are for words left unsaid and = deeds=20 left undone."
 Harriet Beecher Stowe
 
 
----- Original Message -----
From:=20 Beverly=20 Barton
To: Cytometry Mailing = List=20
Sent: Tuesday, May 09, 2006 = 1:59 PM
Subject: Re: Magnetic cell = sorting

Hah, you are lucky, for mouse the yield is more like = 10% and=20 that's what Miltenyi tells you to expect with their system!

Sad = but=20 true.....

Beverly

Beverly E. Barton, Ph.D.
Assistant=20 Professor
Department of Surgery/Division of Urology
UMDNJ-NJMS = MSB=20 G519
185 S. Orange Avenue
Newark, New Jersey = 07103

Telephone=20 973-972-0662
E-mail bartonbe@umdnj.edu
Telefacsimil= e=20 973-972-3892




On May 6, 2006, at 3:50 AM, RAJA = ALAMELU=20 wrote:

Dear Flow list members,
 
My question is not = directly=20 related to flow cytometry, but now and then I have seen questions in = other=20 areas also pop up.
 
We are attempting to do CD4 and CD8 = cells=20 positive isolation from human PBMC, followed by Detach-a bead=20 step, using Dynal magnetic beads. We have followed the = manufacturers=20 instructions as it, is as well as varied each of the conditions for=20 optimization.
 
While the purity and viability after = separation=20 seem to be good, the final cell yield never crosses ~30% of the = total=20 available CD4/CD8 in PBMC. Also the initial input number, magnet = bound cells=20 and unbound cells do not tally.
 
Is 25-30% the = commonly=20 expected yield using this technique? If anybody has observed better = yield,=20 purity and viability, can you please share the=20 procedure?
 
Thanking you in advance,
Alamelu = Raja

Talk=20 is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. = Great rates starting at=20 1=A2/min.
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