12 The average human body has in the order of 10 circulating platelets. They are crucial for hemostasis, and yet excessive platelet activation is a major cause of m- bidity and mortality in western societies. It is therefore not surprising that platelets have become one of the most extensively investigated biological cell types. We are, however, far from understanding precisely how platelets become activated under physiological and pathophysiological conditions. In addition, there are large gaps in our knowledge of platelet production from their giant precursor cell, the megakar- cyte. Understanding megakaryocyte biology will be crucial for the development of platelet gene targeting. The aim of Platelets and Megakaryocytes is therefore to bring together established and recently developed techniques to provide a comprehensive guide to the study of both the platelet and the megakaryocyte. It consists of five s- tions split between two volumes. The more functional assays appear in Volume 1, whereas Volume 2 includes signaling techniques, postgenomic methods, and a n- ber of key perspectives chapters. Part I of Volume 1, Platelets and Megakaryocytes: Functional Assays, describes many well established approaches to the study of platelet function, including aggregometry, secretion, arachidonic acid metabolism, procoagulant responses, pla- let adhesion under static or flow conditions, flow cytometry, and production of microparticles. Although one would ideally wish to perform experiments with human platelets, studies within the circulation using intravital microscopy require the use of animal models, which are described in Chapter 16, vol. 1.

Part I. Platelet Functional Assays Effects of Anticoagulants Used During Blood Collection on Human Platelet Function Jane A. May and Stanley Heptinstall Preparation of Washed Platelet Suspensions From Human and Rodent Blood Jean-Pierre Cazenave, Philippe Ohlmann, Dominique Cassel, Anita Eckly, Beatrice Hechler, and Christian Gachet Platelet Counting Paul Harrison, Carol Briggs, and Samuel J. Machin Electron Microscopy Methods for Studying Platelet Structure and Function James G. White Platelet Aggregation: Turbidimetric Measurements Gavin E. Jarvis Platelet Aggregation in Whole Blood: Impedance and Particle Counting Methods Gavin E. Jarvis Secretion From Dense Granules: Luminescence Method for Adenine Nucleotides M. Fred Heath Platelet Dense-Granule Secretion: The [3H]-5-HT Secretion Assay David Crosby and Alastair W. Poole Analysis of Releasable Nucleotides of Platelets Joachim Jankowski, Lars Henning, and Hartmut Schlüter Studies of Secretion Using Permeabilized Platelets Tara W. Rutledge and Sidney W. Whiteheart Measurement of Platelet Arachidonic Acid Metabolism Richard W. Farndale, Philip G. Hargreaves, Joanna L. Dietrich, and Rosemary J. Keogh Measurement of the Platelet Procoagulant Response Johan W. M. Heemskerk, Paul Comfurius, Marion A. H. Feijge, and Edouard M. Bevers Platelet Adhesion Assays Performed Under Static Conditions Joanne M. Stevens Platelet Adhesion Assays Under Flow Using Matrix Protein-Coupled Adhesion Columns Renata Polanowska-Grabowska and Adrian R. L. Gear Techniques to Examine Platelet Adhesive Interactions Under Flow Suhasini Kulkarni, Warwick S. Nesbitt, Sacha M. Dopheide, Sascha C. Hughan, Ian S. Harper, and Shaun P. Jackson In Vivo Models of Platelet Function and Thrombosis: Study of Real-Time Thrombus Formation Shahrokh Falati, Peter L. Gross, Glenn Merrill-Skoloff, Derek Sim, Robert Flaumenhaft, Alessandro Celi, Barbara C. Furie, and Bruce Furie Adhesion Between Platelets and Leukocytes or Endothelial Cells Gerard B. Nash In Vitro Measurement of High-Shear Platelet Adhesion and Aggregation by the PFA-100 Paul Harrison Flow-Cytometric Analysis of Platelet-Membrane Glycoprotein Expression and Platelet Activation Alison H. Goodall and Jackie Appleby Flow-Cytometric Analysis of Mouse Platelet Function Bernhard Nieswandt, Valerie Schulte, and Wolfgang Bergmeier Platelet Microparticles Shosaku Nomura and Shirou Fukuhara Part II. Megakaryocyte Functional Assays Isolation of Primary Megakaryocytes and Studies of Proplatelet Formation Robert M. Leven Isolation and Culture of Megakaryocyte Precursors Najet Debili, Fawzia Louache, and William Vainchenker Assays of Megakaryocyte Development: Surface Antigen Expression, Ploidy, and Size Anthony Mathur, Ying Hong, Guosu Wang, and Jorge D. Erusalimsky Assays of Megakaryocyte Development: Cytoplasm, Storage Granules, and Demarcation Membranes Arnaud Drouin, Gulie Alimardani, and Elisabeth M. Cramer Thrombopoietin Bioassay Warren S. Alexander and Craig Hyland Culture of Megakaryocytic Cell Lines: Uses and Limitations Norio Komatsu Index