The main aim of this fourth volume is to complement the first three volumes published in 2004 and 2012 by adding advanced methodologies and perspectives. Chapters guide readers through new techniques into the study of platelets and megakaryocytes, including new imaging approaches, new methods for platelet production in vitro, and systems biology approaches. Written in the highly successful Methods in Molecular Biology series format, methods chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and vital tips on troubleshooting or avoiding known pitfalls. Authoritative and up-to-date, Platelets and Megakaryocytes: Volume 4, Additional Protocols and Perspectives adds a wealth of new expertise for the labs of scientists working in this key biological area of study.

For both volumes:
A comprehensive collection of cutting-edge methods for the molecular and cellular analysis of platelets and megakaryocytes. Volume 1 details basic and advanced techniques for analyzing platelet and megakaryocyte function. The approaches presented for platelet analysis include aggregometry, secretion, arachidonic acid metabolism, procoagulent response, platelet adhesion under static and flow conditions, flow cytometry, and production of microparticles. The techniques for study of megakaryocyte function allow the measurement of specialized structures for platelet production, the appearance of platelet-specific surface receptors, and the increase in ploidy. Volume 2 offers techniques for studying many aspects of signaling in platelets and megakaryocytes, as well as state-of-the-art commentaries on our understanding of these cells. The methods include ligand-binding assays, the study of protein and lipid kinases and phosphatases, the analysis of lipid rafts, and the measurement of intracellular calcium levels. There are also techniques for study of electrophysiological events, nitric oxide signaling, venom proteins, and for the internalization of proteins into platelets through permeabilization. This volume also describes the application of molecular and post-genomic methods are to the study of platelets and megakaryocytes. All protocols follow the successful Methods in Molecular Biologya"[ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. The two volumes offer novice and experienced cellbiologists, hematologists, and clinicians not only a benchmark survey of the field, but also a comprehensive library of proven techniques essential for productive research on platelets and megakaryocytes today.

For both volumes:
A comprehensive collection of cutting-edge methods for the molecular and cellular analysis of platelets and megakaryocytes. Volume 1 details basic and advanced techniques for analyzing platelet and megakaryocyte function. The approaches presented for platelet analysis include aggregometry, secretion, arachidonic acid metabolism, procoagulent response, platelet adhesion under static and flow conditions, flow cytometry, and production of microparticles. The techniques for study of megakaryocyte function allow the measurement of specialized structures for platelet production, the appearance of platelet-specific surface receptors, and the increase in ploidy. Volume 2 offers techniques for studying many aspects of signaling in platelets and megakaryocytes, as well as state-of-the-art commentaries on our understanding of these cells. The methods include ligand-binding assays, the study of protein and lipid kinases and phosphatases, the analysis of lipid rafts, and the measurement of intracellular calcium levels. There are also techniques for study of electrophysiological events, nitric oxide signaling, venom proteins, and for the internalization of proteins into platelets through permeabilization. This volume also describes the application of molecular and post-genomic methods for the study of platelets and megakaryocytes. All protocols follow the successful Methods in Molecular Biologya"[ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. The two volumes offer novice and experienced cellbiologists, hematologists, and clinicians not only a benchmark survey of the field, but also a comprehensive library of proven techniques essential for productive research on platelets and megakaryocytes today.

New techniques to study cell signaling and function can develop at a staggering pace; however, many approaches are as valid today as on the day they were established. Thus, the main aim of Platelets and Megakaryocytes: Volume 3, Additional Protocols and Perspectives is to complement the first two volumes published in 2004 by adding recently developed state of the art techniques. Conveniently divided into three sections, this detailed volume covers techniques to study platelet function, approaches to investigate megakaryocyte function, and perspectives on important overall concepts in the field of megakaryocyte and platelet biology. Written in the highly successful Methods in Molecular Biology (TM) series format, methods chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and vital tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Platelets and Megakaryocytes: Volume 3, Additional Protocols and Perspectives adds a wealth of new expertise for the labs of scientists working in this key biological area of study.

New techniques to study cell signaling and function can develop at a staggering pace; however, many approaches are as valid today as on the day they were established. Thus, the main aim of Platelets and Megakaryocytes: Volume 3, Additional Protocols and Perspectives is to complement the first two volumes published in 2004 by adding recently developed state of the art techniques. Conveniently divided into three sections, this detailed volume covers techniques to study platelet function, approaches to investigate megakaryocyte function, and perspectives on important overall concepts in the field of megakaryocyte and platelet biology. Written in the highly successful Methods in Molecular Biology (TM) series format, methods chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and vital tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Platelets and Megakaryocytes: Volume 3, Additional Protocols and Perspectives adds a wealth of new expertise for the labs of scientists working in this key biological area of study.

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.

12 The average human body has on 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.