Main description:

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.

Contents:

1. Immobilisation of Non-activated Unfixed Platelets for Real Time Single-cell Analysis

Alexander P Bye, Zeki Ilkan, Amanda J Unsworth, and Chris I Jones

2. Imaging Platelets and Megakaryocytes by High Resolution Laser Fluorescence Microscopy

Fred G. Pluthero and Walter H. A. Kahr

3. Single Molecule Localisation and Structured Illumination Microscopy of Platelet Proteins

Natalie S. Poulter, Abdullah O. Khan , Chiara Pallini, and Steven G. Thomas

4. Electron Tomography and Correlative Approaches in Platelet Studies
Kasia B. Engberts, Cor Seinen, Willie J.C. Geerts, and Harry F.G. Heijnen

5. Screening and High-throughput Platelets Assays

Alexander P Bye, Amanda J Unsworth, and Jonathan M Gibbins

6. High-throughput Signaling Profiling in Blood Platelets by Multiplexed Phosphoflow Cytometry

Benjamin E.J. Spurgeon and Khalid M. Naseem

7. Precise Quantification of Platelet Proteins and their Phosphorylation States

Francoise Mazet and Michael J. Fry

8. The Study of Platelet Receptors using Artificial Lipid Bilayers

Michael L. Dustin and Alice Y. Pollitt

9. Three Dimensional Culture in a Methylcellulose-based Hydrogel to Study the Impact of Stiffness on Megakaryocyte Differentiation

Alicia Aguilar, Julie Boscher, Fabien Pertuy, Christian Gachet, and Catherine Leon

10. Differentiation of Human Pluripotent Stem Cells to Megakaryocytes by Transcription Factor Driven Forward Programming

Thomas P. Moreau, Amanda L. Evans, and Cedric JG. Ghevaert

11. Three-dimensional Tissue Models for Studying Ex Vivo Megakaryocytopoiesis and Platelet Production

Christian A. Di Buduo, Vittorio Abbonante, Lorenzo Tozzi, David L. Kaplan, and Alessandra Balduini

12. Fluorescence Approaches to Image and Quantify the Demarcation Membrane System in Living Megakaryocytes

Sangar Osman, Daniel Dalmay, and Martyn P. Mahaut-Smith

13. High-resolution 3D Imaging of Megakaryocytes using Focused Ion Beam-Scanning Electron Microscopy

Anita Eckly, Jean-Yves Rinckel, Fabienne Proamer, and Christian Gachet

14. Optical Clearing of Murine Bones to Study Megakaryocytes in Intact Bone Marrow Using Light-sheet Fluorescence Microscopy

Maximilian G. Gorelashvili, Katrin G. Heinze, and David Stegner

15. Mathematical Techniques for Understanding Platelet Regulation and the Development of New Pharmacological Approaches

J.L. Dunster, M.A. Panteleev, J.M. Gibbins, and A.N. Sveshnikova