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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