Re: flow sensitivity

From: Glenn Paradis (gap@mit.edu)
Date: Wed Jan 09 2002 - 13:50:18 EST


Hi Everyone,

I would be interested in other products that would increase
sensitivity for cell surface staining applications such as the one
Dr. Kaplan decribes below.  If you know of one, please email me.  I
will be happy to summarize the replies.

Thank you,

Glenn
MIT


*************************************************
Glenn Paradis
MIT Flow Cytometry Core Facility
E18-463
Massachusetts Institute of Technology
Cambridge, MA 02139
(617)253-6454 lab
(617)452-4132 office
gap@mit.edu
http://web.mit.edu/flowcytometry/www/
*************************************************




>Cytometrists,
>
>I am writing about Richard Haugland's communication from December 22.  In
>response to the low sensitivity of detection in flow cytometry, he wrote
>that he has been working on using Tyramide Signal Amplification (TSA) to
>increase the sensitivity of flow cytometric detection.  He provides a link
>to data that show excellent results with TSA to enhance the detection of
>the EGF receptor.  He also indicates that he has similar data for detection
>of the estrogen receptor.
>
>We agree with Dr. Haugland's enthusiasm for TSA.  We have used a modified
>TSA procedure for the detection of both cell surface and intracellular
>molecules by flow cytometry.  We refer to our technique as enzymatic
>amplification staining (EAS).  The staining that we have obtained has been
>validated via an independent attribute of the molecule being detected
>(Kaplan, D, D Smith.  Enzymatic amplification staining for flow cytometric
>analysis of cell surface molecules.  Cytometry  40:81-85, 2000).  Moreover,
>we have used EAS to provide high resolution immunophenotyping for leukemic
>cells from patients which includes 4-color analysis (Kaplan, D, H Meyerson,
>K Lewandowska.  High resolution immunophenotypic analysis of chronic
>lymphocytic leukemia cells by enzymatic amplification staining (EAS).  Amer
>J. Clin. Path.  116:429-36, 2001) and for an analysis of CD5 expression on
>human B lymphocytes which includes both surface and intracellular staining
>(Kaplan, D, D Smith, H Meyerson, N Pecora, K Lewandowska.  CD5 expression
>by B lymphocytes and its regulation upon Epstein-Barr Virus transformation.
>  Proc. Natl. Acad. Sci. USA.  98:1385-13853, 2001).  We also have 4 other
>manuscripts in press that feature the EAS technology.  Our technology has
>been patented and we have licensed this technology to Flow-Amp Systems,
>Ltd. which offers a variety of kits for performing EAS.  Although these
>kits have only been available for a little more than a year, already 3
>papers have been published by other investigators using our kits
>(J.Exp.Med. 194:1339; 2001; J Immunol 167:6654, 2001; and Lab Invest
>81:1223, 2001).  The kits are available from Flow-Amp Systems.  The
>company's website includes contact information, lots of examples of data,
>protocols, and troubleshooting guides.  The website follows:
>
>http://www.flow-amp.com/
>
>I have also included 3 attachments demonstrating the power of EAS.
>
>David
>
>David Kaplan, MD, PhD
>Co-Founder, Flow-Amp Systems, Ltd.
>11000 Cedar Avenue
>Cleveland, OH  44106
>tel:  (216)721-0590
>office tel:  (216)368-1279
>email:  dkaplan@flow-amp.com
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--
Glenn Paradis



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