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Platelets and Megakaryocytes

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Cover of 'Platelets and Megakaryocytes'

Table of Contents

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    Book Overview
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    Chapter 1 Immobilization of Nonactivated Unfixed Platelets for Real-Time Single-Cell Analysis
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    Chapter 2 Imaging Platelets and Megakaryocytes by High-Resolution Laser Fluorescence Microscopy
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    Chapter 3 Single-Molecule Localization and Structured Illumination Microscopy of Platelet Proteins
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    Chapter 4 Electron Tomography and Correlative Approaches in Platelet Studies
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    Chapter 5 Screening and High-Throughput Platelet Assays
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    Chapter 6 High-Throughput Signaling Profiling in Blood Platelets by Multiplexed Phosphoflow Cytometry
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    Chapter 7 Precise Quantification of Platelet Proteins and Their Phosphorylation States
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    Chapter 8 The Study of Platelet Receptors Using Artificial Lipid Bilayers
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    Chapter 9 Three-Dimensional Culture in a Methylcellulose-Based Hydrogel to Study the Impact of Stiffness on Megakaryocyte Differentiation
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    Chapter 10 Differentiation of Human Pluripotent Stem Cells to Megakaryocytes by Transcription Factor-Driven Forward Programming
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    Chapter 11 Three-Dimensional Tissue Models for Studying Ex Vivo Megakaryocytopoiesis and Platelet Production
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    Chapter 12 Fluorescence Approaches to Image and Quantify the Demarcation Membrane System in Living Megakaryocytes
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    Chapter 13 High-Resolution 3D Imaging of Megakaryocytes Using Focused Ion Beam-Scanning Electron Microscopy
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    Chapter 14 Optical Clearing of Murine Bones to Study Megakaryocytes in Intact Bone Marrow Using Light-Sheet Fluorescence Microscopy
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    Chapter 15 Mathematical Techniques for Understanding Platelet Regulation and the Development of New Pharmacological Approaches
Attention for Chapter 5: Screening and High-Throughput Platelet Assays
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Chapter title
Screening and High-Throughput Platelet Assays
Chapter number 5
Book title
Platelets and Megakaryocytes
Published in
Methods in molecular biology, September 2018
DOI 10.1007/978-1-4939-8585-2_5
Pubmed ID
Book ISBNs
978-1-4939-8584-5, 978-1-4939-8585-2
Authors

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

Abstract

High-throughput assays are important biological research tools but are rarely utilized for platelet research. However, screening compounds for efficacy against a physiologically relevant cellular response in primary cells such as platelets can be an advantageous approach to compound screening and drug development. In this section we describe a panel of three high-throughput microtiter plate assays designed for platelets that can be used as the basis for compound screening, or be modified and used individually to increase throughput in platelet research laboratories. The platelet adhesion assay has the lowest requirement for platelet numbers and is therefore capable of the greatest throughput and so is suggested as the primary screen used to identify hits. A secondary screen against the "gold standard" of platelet function, aggregation, is used to confirm and further characterize hits. Finally, a Ca2+ assay is used for initial mechanistic characterization to begin the process of elucidating the mode of action of hit compounds.