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.
This book details methods for the selection of key cellular, molecular and biochemical techniques that are used in studying the many and varied functions of microglia. It features step-by-step, readily reproducible laboratory protocols.
Features tips on trouble shooting and avoiding known pitfalls
Part I: Overview of Microglia Biology1. A Brief Overview of Multi-Talented MicrogliaBertrand Joseph and Jose Luis Venero Part II: Isolation and Culture of Microglia2. Cell Culturing of Human and Murine Microglia Cell LinesJohanna Rodhe 3. Microglia Isolation from Adult Mouse Brain Jae-Kyung Lee, Malú G Tansey 4. Preparation of Primary Microglia Cultures from Postnatal Mouse and Rat BrainsTomas Deierborg 5. Isolation of Murine Postnatal Brain Microglia for Phenotypic Characterization Using Magnetic Cell Separation TechnologyAshley S. Harms and Malú G. Tansey 6. Isolation and Culture of Adult Human Microglia within Mixed Glial Cultures for Functional Experimentation and High Content AnalysisAmy M. Smith, Hannah M. Gibbons, Claire Lill, Richard L. M. Faull and Mike Dragunow Part III: Depletion and Transduction of Microglia7. Depletion of Microglia from Primary Cellular CulturesLorena Pont-Lezica, Sabrina Colasse and Alain Bessis 8. Lentiviral Transduction of Cultured MicrogliaTakahiro Masuda, Makoto Tsuda, Hidetoshi Tozaki-Saitoh and Kazuhide Inoue Part IV: Analysis of Microglial Cytokine Production9. Microglial Activation: Measurement of Cytokines by Flow CytometryDeepak Kumar Kaushik and AnirbanBasu 10. In situ Hybridization of Cytokine mRNA using Alkaline Phosphatase Labelled Oligodeoxynucleotide ProbesBettina Clausen, Christina Fenger and Bente Finsen 11. Use of Meso-scale DiscoveryTM to Examine Cytokinecontentin Microglia Cell SupernatantMiguel A. Burguillos Part V: Analysis of Microglia Activation12. Analysis of Microglial Production of Reactive Oxygen and Nitrogen SpeciesUrteNeniskyte and Guy C. Brown 13. Quantification of Active Caspase-3 and Active Caspase-8 in Microglia CellsEdel Kavanagh 14. Quantification of Microglial Phagocytosis by a Flow Cytometer-based AssayRefik Pul, Chittappen Kandiyil Prajeeth, Martin Stangel 15. Quantification of Microglial Proliferation and Apoptosis by Flow Cytometry Alicia A. Babcock, Martin Wirenfeldt, and Bente Finsen 16. Fluorescence Imaging of Intracellular Ca2+, Na+ and H+ in Cultured Microglia Tom Schilling, and Claudia Eder 17. Patch Clamp Protocols to Study Ion Channel Activity in Microglia Tom Schilling, and Claudia Eder Part VI: Analysis of Microglia Polarization18. Studying M1 and M2 States in Adult MicrogliaSadanand M. Gaikwad and Michael T. Heneka 19. Isolating, Culturing, and Polarizing Primary Human Adult and Fetal MicrogliaBryce A. Durafourt, Craig S. Moore, Manon Blain, and Jack P. Antel Part VII: Co-culture Systems to Analysis Microglia Interactions with Other Cell Types20. Understanding Microglia-neuron Cross-talk: Relevance of the Microglia-neuron Co-culturesFernando G Correa, Miriam Hernangómez, and Carmen Guaza 21. Preparation of Rodent Primary Cultures for Neuron-Glia, Mixed Glia, Enriched Microglia, and Reconstituted Cultures with MicrogliaShih-Heng Chen, Esteban A. Oyarzabal and Jau-Shyong Hong Part VIII: Analysis of Microglia Functions in vivo22. Microglia Detection by Enzymatic HistochemistryBeatriz Almolda, Berta González and Bernardo Castellano 23. Tomato Lectin Histochemistry for Microglial VisulizationNàdia Villacampa, Beatriz Almolda, Berta González and Bernardo Castellano 24. Immunohistochemical Detection of MicrogliaRocío M. de Pablos, Ana M. Espinosa-Oliva, and Antonio J. Herrera 25. Intrathecal Infusion of Microglia CellsTakahiro Masuda, Makoto Tsuda, Hidetoshi Tozaki-Saitoh and Kazuhide Inoue 26. Intracranial Injection of LPS in Rat as Animal Model of NeuroinflammationAna M. Espinosa-Oliva, Rocío M. de Pablos, and Antonio J. Herrera 27. Analyses of Microglia Effector Function Using CX3CR1-GFP Knock In MiceJenny A. Garcia, Sandra M. Cardona and Astrid E. Cardona 28. In vivo Two-photon Microscopy of MicrogliaSatoru Kondo and Shigeo Okabe 29. Use of Confocal Microscopy in the Study of Microglia in a Brain Metastasis ModelManuel Sarmiento