Full Description
Key discoveries concerning the different biological functions of microglia in health and disease have attracted scientists from various fields. In Microglia: Methods and Protocols, expert researchers in the field detail methods for selection of the key cellular, molecular and biochemical techniques that are used in studying the many and varied functions of this fascinating cell. These methods and techniques include microglia cell culture for studying microglia activation and functions, as well as their interaction with other cell types both in vitro and in vivo. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Microglia: Methods and Protocols is a useful resource for cell biologists, molecular biologists, immunologists, oncologist and neuroscientists.
Contents
A Brief Overview of Multi-Talented Microglia.- Cell Culturing of Human and Murine Microglia Cell Lines.- Microglia Isolation from Adult Mouse Brain.- Preparation of Primary Microglia Cultures from Postnatal Mouse and Rat Brains.- Isolation of Murine Postnatal Brain Microglia for Phenotypic Characterization Using Magnetic Cell Separation Technology.- Isolation and Culture of Adult Human Microglia within Mixed Glial Cultures for Functional Experimentation and High Content Analysis.- Depletion of Microglia from Primary Cellular Cultures.- Lentiviral Transduction of Cultured Microglia.- Microglial Activation: Measurement of Cytokines by Flow Cytometry.- In situ Hybridization of Cytokine mRNA using Alkaline Phosphatase Labelled Oligodeoxynucleotide Probes.- Use of Meso-scale DiscoveryTM to Examine Cytokinecontentin Microglia Cell Supernatant.- Analysis of Microglial Production of Reactive Oxygen and Nitrogen Species.- Quantification of Active Caspase-3 and Active Caspase-8 in Microglia Cells.- Quantification of Microglial Phagocytosis by a Flow Cytometer-based Assay.- Quantification of Microglial Proliferation and Apoptosis by Flow Cytometry.- Fluorescence Imaging of Intracellular Ca2+, Na+ and H+ in Cultured Microglia.- Patch Clamp Protocols to Study Ion Channel Activity in Microglia.- Studying M1 and M2 States in Adult Microglia.- Isolating, Culturing, and Polarizing Primary Human Adult and Fetal Microglia.- Understanding Microglia-neuron Cross-talk: Relevance of the Microglia-neuron Co-cultures.- Preparation of Rodent Primary Cultures for Neuron-Glia, Mixed Glia, Enriched Microglia, and Reconstituted Cultures with Microglia.- Microglia Detection by Enzymatic Histochemistry.- Tomato Lectin Histochemistry for Microglial Visulization.- Immunohistochemical Detection of Microglia.- Intrathecal Infusion of Microglia Cells.- Intracranial Injection of LPS in Rat as Animal Model of Neuroinflammation.- Analyses ofMicroglia Effector Function Using CX3CR1-GFP Knock In Mice.- In vivo Two-photon Microscopy of Microglia.- Use of Confocal Microscopy in the Study of Microglia in a Brain Metastasis Model.
