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 <0IZB00GIGWFG9E@pop.mtnl.net.in> for modi@mtnl.net.in; Tue, 16 May 2006 04:12:52 +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 <0IZB00CGBWFGLG@mx.mtnl.net.in> for modi@mtnl.net.in (ORCPT modi@mtnl.net.in); Tue, 16 May 2006 04:12:52 +0530 (IST) Received: from flowcyt.cyto.purdue.edu (128.210.61.51) by mtnl-av (202.159.228.19); Tue, 16 May 2006 04:03:29 +0530 Received: (from root@localhost) by flowcyt.cyto.purdue.edu (8.12.10/8.12.10) id k4FGpfDU094608 for cyto-sendout; Mon, 15 May 2006 12:51:41 -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 k4F7UmQ4087015 for ; Mon, 15 May 2006 02:30:48 -0500 Received: (from mailnull@localhost) by flowcyt.cyto.purdue.edu (8.12.10/8.12.10/Submit) id k4F7UmJA087014 for cytolist; Mon, 15 May 2006 03:30:48 -0400 (EDT envelope-from mailnull) Received: from plexus.uke.uni-hamburg.de (plexus.uke.uni-hamburg.de [134.100.96.1]) by flowcyt.cyto.purdue.edu (8.12.10/8.12.10) with ESMTP id k4F7Ui7s087008 for ; Mon, 15 May 2006 03:30:44 -0400 Received: from WORKSTATION2 ([134.100.96.116]) by plexus.uke.uni-hamburg.de (8.12.10/8.12.10/SuSE Linux 0.7) with SMTP id k4F7UevQ011220; Mon, 15 May 2006 09:30:41 +0200 Content-return: prohibited Date: Mon, 15 May 2006 09:28:28 +0200 From: Boris Fehse Subject: AW: Magnetic cell sorting In-reply-to: <000001c67533$21f90e70$7101a8c0@VALUEDD9017A01> To: cyto-inbox Message-id: MIME-version: 1.0 X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2910.0) Content-type: multipart/mixed; boundary="Boundary_(ID_cXx4zKVyAFajjsKtjow5Mg)" Importance: Normal X-Priority: 3 X-MSMail-priority: Normal X-eSCM-MailFrom: X-Spam-Score: -4.487 () BAYES_00,HTML_60_70,HTML_FONTCOLOR_BLUE,HTML_FONTCOLOR_UNKNOWN,HTML_MESSAGE X-Scanned-By: MIMEDefang 2.39 X-PMFLAGS: 570950016 0 1 87020.cnm X-Antivirus: AVG for E-mail 7.1.392 [268.5.6/339] Original-recipient: rfc822;modi@mtnl.net.in This is a multi-part message in MIME format. --Boundary_(ID_cXx4zKVyAFajjsKtjow5Mg) Content-type: multipart/alternative; boundary="Boundary_(ID_wDx/Rhm3OtHMwpgt3x2Xbw)" --Boundary_(ID_wDx/Rhm3OtHMwpgt3x2Xbw) X-MIME-Autoconverted: from 8bit to quoted-printable by flowcyt.cyto.purdue.edu id k4FGpfDV094608 Content-type: text/plain; charset=iso-8859-1 Content-transfer-encoding: quoted-printable RE: Magnetic cell sortingDear (MAC) sorters, I hope we won't be confronted with a series of ads and sarcastic letters from different companies involved in magnetic cell sorting. Obviously all systems have their +es and -es. (We have tried quite a number of them...) Best regards Boris p.s. Sorry for that, Jenny - I checked our data and in our hands it's no problem to sort CD4 and CD8 with high purity (>90%) with Miltenyi beads. (I'm not with the company, and have no any commercial interest). ...;) B. Boris Fehse, PhD University Medical Center Hamburg-Eppendorf -----Urspr=FCngliche Nachricht----- Von: Jenny Cabanillas [mailto:cabanillas@optonline.net] Gesendet: Donnerstag, 11. Mai 2006 21:43 An: Cytometry Mailing List Betreff: FW: Magnetic cell sorting Dear Roger, I don=92t believe we had the pleasure of meeting. I am part of the Invitrogen CSA team, and I=92m responsible for the Dynal Beads in NJ/NY. It amazing that you would say 90% purity and recovery since from my canvassing I have heard from researchers that the results vary incredibly from Experiment to experiment (that=92s if the columns don=92t clog of cours= e). It varies from applications but over 90% is a bit high. I would check my dat= a if I were you J Proof is in the pudding! Having fun selling Dynal beads, Jenny Cabanillas-McCauley -----Original Message----- From: Roger Burger [mailto:rburger@miltenyibiotec.com] Sent: Tuesday, May 09, 2006 5:44 PM To: Cytometry Mailing List Subject: RE: Magnetic cell sorting Hello Beverly I hope you meant to say, "Hah, you are lucky, for mouse the yield is more like 10% and that's wh= at Dynal tells you to expect with their system!" With the Miltenyi System and MACS=AE CD4 and CD8 Microbeads you can achieve >90% purity and >90% recovery. Regards, Roger A. Burger, Ph.D. Technical Marketing Scientist Miltenyi Biotec, Inc. 12740 Earhart Ave. Auburn, CA 95602 Phone: 530-887-5351 Fax: 530-745-2851 www.miltenyibiotec.com -----Original Message----- From: Beverly Barton [mailto:bartonbe@umdnj.edu] Sent: Tuesday, May 09, 2006 5:00 AM To: Cytometry Mailing List Subject: Re: Magnetic cell sorting Hah, you are lucky, for mouse the yield is more like 10% and that's wha= t 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, My question is not directly related to flow cytometry, but now and then= I have seen questions in other areas also pop up. 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. While the purity and viability after separation seem to be good, the fi= nal cell yield never crosses ~30% of the total available CD4/CD8 in PBMC. Als= o the initial input number, magnet bound cells and unbound cells do not tal= ly. 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? Thanking you in advance, Alamelu Raja Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great ra= tes starting at 1=A2/min. --Boundary_(ID_wDx/Rhm3OtHMwpgt3x2Xbw) Content-type: text/html; charset=iso-8859-1 Content-transfer-encoding: quoted-printable RE: Magnetic cell = sorting
Dear=20 (MAC) sorters,
 
I=20 hope we won't be confronted with a series of ads and sarcastic = letters from=20 different companies involved in magnetic cell sorting. Obviously all = systems=20 have their +es and -es. (We have tried quite a number of=20 them...)
 
Best=20 regards
Boris
 
p.s.=20 Sorry for that, Jenny - I checked our data and in our hands it's no = problem to=20 sort CD4 and CD8 with high purity (>90%) with Miltenyi beads. (I'm = not with=20 the company, and have no any commercial interest). ...;) = B.
 
 
Boris=20 Fehse, PhD
University Medical Center Hamburg-Eppendorf
 
 
 
 
 
 
 
-----Urspr=FCngliche Nachricht-----
Von: Jenny = Cabanillas=20 [mailto:cabanillas@optonline.net]
Gesendet: Donnerstag, 11. = Mai 2006=20 21:43
An: Cytometry Mailing List
Betreff: FW: = Magnetic=20 cell sorting

Dear=20 Roger,

I don=92t = believe we=20 had the pleasure of meeting.  = I am=20 part of the Invitrogen CSA team, and I=92m responsible for the Dynal = Beads in=20 NJ/NY.

It amazing = that you=20 would say 90% purity and recovery since from my canvassing I have = heard from=20 researchers that the results vary incredibly = from

Experiment = to=20 experiment (that=92s if the columns don=92t clog of=20 course).  It varies from applications = but over=20 90% is a bit high. I would check my data if I were you = J

 

Proof is in = the=20 pudding! Having fun selling Dynal beads,

Jenny=20 Cabanillas-McCauley

 

 

 

 

-----Original=20 Message-----
From: = Roger=20 Burger [mailto:rburger@miltenyibiotec.com]
Sent: Tuesday, May 09, 2006 = 5:44=20 PM
To: Cytometry = Mailing=20 List
Subject: RE: = Magnetic=20 cell sorting

 

Hello = Beverly=20

I=20 hope you meant to say,

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

 With the Miltenyi = System and=20 MACS=AE  CD4 and CD8 Microbeads you can achieve >90% purity = and >90%=20 recovery. 

Regards,=20

 

Roger A. Burger, Ph.D.=20
Technical Marketing = Scientist=20
Miltenyi Biotec, Inc.=20
12740 Earhart Ave.=20
Auburn, CA 95602=20
Phone: 530-887-5351=20
Fax: 530-745-2851=20
www.miltenyibiotec.com=20

-----Original=20 Message-----
From: Beverly Barton=20 [mailto:bartonbe@umdnj.edu]=20
Sent: Tuesday, May 09, = 2006 5:00=20 AM
To: Cytometry Mailing=20 List
Subject: Re: Magnetic = cell=20 sorting

 

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

 

Sad but true.....=20

 

Beverly=20

 

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

 

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


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

 

Dear Flow list members,=20
  =
My=20 question is not directly related to flow cytometry, but now and then I = have=20 seen questions in other areas also pop up. =


We are=20 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=20 the manufacturers instructions as it, is as well as varied each of the = conditions for optimization.


While the=20 purity and viability after separation seem to be good, the final cell = yield=20 never crosses ~30% of the total available CD4/CD8 in PBMC. Also the = initial=20 input number, magnet bound cells and unbound cells do not tally.=20


Is 25-30%=20 the commonly expected yield using this technique? If anybody has = observed=20 better yield, purity and viability, can you please share the = procedure?=20


Thanking=20 you in advance,
Alamelu Raja=20

 

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

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