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Prepublished online as a Blood First Edition Paper on June 21, 2002; DOI 10.1182/blood-2002-03-0902. This Article Full Text (PDF) All Versions of this Article: 2002-03-0902v1 100/8/2787    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Balasubramanian, V. Articles by Nemerson, Y. Search for Related Content PubMed PubMed Citation Articles by Balasubramanian, V. Articles by Nemerson, Y. Social Bookmarking                   What's this? Submitted March 22, 2002 Accepted May 15, 2002 Platelets, circulating tissue factor, and fibrin colocalize in ex-vivo thrombi: Real-time fluorescence images of thrombus formation and propagation under defined flow conditions Viji Balasubramanian, Eric Grabowski, Alessandra Bini, and Yale Nemerson* Thrombosis Research, Mount Sinai School of Medicine, New York, NY, USA Cardiovascular Thrombosis Laboratory, Massachusetts General Hospital-Harvard Medical School, Boston, MA, USA New York Blood Center and Department of Pathology, Columbia University College of Physicians and Surgeons, New York, New York, NY, USA * Corresponding author; email: Yale.Nemerson{at}mssm.edu. While it is generally accepted that the initial event in coagulation and intravascular thrombus formation is the exposure of tissue factor (TF) to blood, there is still little agreement about the mechanisms of thrombus propagation and the identities of the molecular species participating in this process. In this study, we characterized the thrombotic process in real-time and under defined flow conditions, in order to determine the relative contribution and spatial distribution of three components of the thrombi: circulating or blood-borne TF (cTF), fibrin, and platelets. For this purpose, we utilized high-sensitivity, multi-color immunofluorescence microscopy coupled with a laminar flow chamber. Freshly drawn blood, labeled with mepacrine (marker for platelets and white cells), anti-hTF1Alexa 568(marker for tissue factor), and anti-T2G1Cy-5 (marker for fibrin) was perfused over collagen-coated glass slides at wall shear rates of 100 and 650 s-1. A motorized filter cube selector facilitated imaging every 5 seconds at one of 3 different wavelengths, corresponding to optimal wavelengths for the 3 markers above. Real-time video recordings obtained during 10 discrete experiments show rapid deposition of platelets and fibrin onto collagen-coated glass. Overlay images of platelets, fibrin, and cTF clearly demonstrate co-localization of these three components in growing thrombi. These data further support our earlier observations that, in addition to TF present in the vessel wall, there is a pool of TF in circulating blood that contributes to the propagation of thrombosis at a site of vascular injury. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: T. Orfeo, K. E. Brummel-Ziedins, M. Gissel, S. Butenas, and K. G. Mann The Nature of the Stable Blood Clot Procoagulant Activities J. Biol. Chem., April 11, 2008; 283(15): 9776 - 9786. [Abstract] [Full Text] [PDF] A. T. Nurden Platelets and Tissue Remodeling: Extending the Role of the Blood Clotting System Endocrinology, July 1, 2007; 148(7): 3053 - 3055. [Full Text] [PDF] J. J. Hathcock Flow Effects on Coagulation and Thrombosis Arterioscler Thromb Vasc Biol, August 1, 2006; 26(8): 1729 - 1737. 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