Microglia : methods and protocols

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. Part I Overview Of Microglia Biology -- Brief Overview Of Multitalented Microglia / Bertrand Joseph And José Luis Venero -- Part Ii Isolation And Culture Of Microglia -- Cell Culturing Of Human And Murine Microglia Cell Lines / Johanna Rodhe -- Microglia Isolation From Adult Mouse Brain / Jae-kyung Lee And Malú G. Tansey -- Preparation Of Primary Microglia Cultures From Postnatal Mouse And Rat Brains / Tomas Deierborg -- Isolation Of Murine Postnatal Brain Microglia For Phenotypic Characterization Using Magnetic Cell Separation Technology / Ashley S. Harms And Malú G. Tansey -- Isolation And Culture Of Adult Human Microglia Within Mixed Glial Cultures For Functional Experimentation And High-content Analysis / Amy M. Smith [and Others] -- Part Iii Depletion And Transduction Of Microglia -- Depletion Of Microglia From Primary Cellular Cultures / Lorena Pont-lezica, Sabrina Colasse, And Alain Bessis -- Lentiviral Transduction Of Cultured Microglia / Takahiro Masuda [and Others] -- Part Iv Analysis Of Microglial Cytokine Production -- Microglial Activation : Measurement Of Cytokines By Flow Cytometry / Deepak Kumar Kaushik And Anirban Basu -- In Situ Hybridization Of Cytokine Mrna Using Alkaline Phosphatase-labelled Oligodeoxynucleotide Probes / Bettina Clausen, Christina Fenger, And Bente Finsen -- Use Of Meso-scale Discovery To Examine Cytokine Content In Microglia Cell Supernatant / Miguel A. Burguillos -- Part V Analysis Of Microglia Activation -- Analysis Of Microglial Production Of Reactive Oxygen And Nitrogen Species / Urte Neniskyte And Guy C. Brown -- Quantification Of Active Caspase-3 And Active Caspase-8 In Microglia Cells / Edel Kavanagh -- Quantification Of Microglial Phagocytosis By A Flow Cytometer-based Assay / Refik Pul, Kandiyil Prajeeth Chittappen, And Martin Stangel -- Quantification Of Microglial Proliferation And Apoptosis By Flow Cytometry / Alicia A. Babcock, Martin Wirenfeldt, And Bente Finsen -- Fluorescence Imaging Of Intracellular Ca2+, Na+, And H+ In Cultured Microglia / Tom Schilling And Claudia Eder -- Patch Clamp Protocols To Study Ion Channel Activity In Microglia / Tom Schilling And Claudia Eder -- Part Vi Analysis Of Microglia Polarization -- Studying M1 And M2 States In Adult Microglia / Sadanand M. Gaikwad And Michael T. Heneka -- Isolating, Culturing, And Polarizing Primary Human Adult And Fetal Microglia / Bryce A. Durafourt [and Others] -- Part Vii Co-culture Systems To Analysis Microglia Interactions With Other Cell Types -- Understanding Microglia-neuron Cross Talk : Relevance Of The Microglia-neuron Cocultures / Fernando G. Correa, Miriam Hernangómez, And Carmen Guaza -- Preparation Of Rodent Primary Cultures For Neuron-glia, Mixed Glia, Enriched Microglia, And Reconstituted Cultures With Microglia / Shih-heng Chen, Esteban A. Oyarzabal, And Jau-shyong Hong -- Part Viii Analysis Of Microglia Functions In Vivo -- Microglia Detection By Enzymatic Histochemistry / Beatriz Almolda, Berta González, And Bernardo Castellano -- Tomato Lectin Histochemistry For Microglial Visualization / Nàdia Villacampa [and Others] -- Immunohistochemical Detection Of Microglia / Rocío M. De Pablos, Ana M. Espinosa-oliva, And Antonio J. Herrera -- Intrathecal Infusion Of Microglia Cells / Takahiro Masuda [and Others] -- Intracranial Injection Of Lps In Rat As Animal Model Of Neuroinflammation / Ana M. Espinosa-oliva, Rocío M. De Pablos, And Antonio J. Herrera -- Analyses Of Microglia Effector Function Using Cx3cr1-gfp Knock-in Mice / Jenny A. Garcia, Sandra M. Cardona, And Astrid E. Cardona -- In Vivo Two-photon Microscopy Of Microglia / Satoru Kondo And Shigeo Okabe -- Use Of Confocal Microscopy In The Study Of Microglia In A Brain Metastasis Model / Manuel Sarmiento. Edited By Bertrand Joseph, Department Of Oncology-pathology, Karolinska Institutet, Cancer Centrum Karolinska, Stockholm, Sweden, José Luis Venero, Departamento De Bioquimica Y Biologia Molecular, Facultad De Farmacia, Universidad De Sevilla, Sevilla, Spain, Instituto De Biomedicina De Sevilla, Hospital Universitaria, Sevilla, Spain. Includes Bibliographical References And Index. Part I Overview of microglia biology -- Brief overview of multitalented microglia -- Bertrand Joseph and José Luis Venero -- Part II Isolation and culture of microglia -- Cell culturing of human and murine microglia cell lines -- Johanna Rodhe Microglia isolation from adult mouse brain -- Jae-Kyung Lee and Malú G. Tansey Preparation of primary microglia cultures from postnatal mouse and rat brains -- Tomas Deierborg Isolation of murine postnatal brain microglia for phenotypic characterization using magnetic cell separation technology -- Ashley S. Harms and Malú G. Tansey Isolation and culture of adult human microglia within mixed glial cultures for functional experimentation and high-content analysis -- Amy M. Smith ... [et al.] -- Part III Depletion and transduction of microglia -- Depletion of microglia from primary cellular cultures -- Lorena Pont-Lezica, Sabrina Colasse, and Alain Bessis Lentiviral transduction of cultured microglia -- Takahiro Masuda ... [et al.] -- Part IV Analysis of microglial cytokine production -- Microglial activation : measurement of cytokines by flow cytometry -- Deepak Kumar Kaushik and Anirban Basu In situ hybridization of cytokine mRNA using alkaline phosphatase-labelled oligodeoxynucleotide probes -- Bettina Clausen, Christina Fenger, and Bente Finsen Use of meso-scale discovery to examine cytokine content in microglia cell supernatant -- Miguel A. Burguillos -- Part V Analysis of microglia activation -- Analysis of microglial production of reactive oxygen and nitrogen species -- Urte Neniskyte and Guy C. Brown Quantification of active caspase-3 and active caspase-8 in microglia cells -- Edel Kavanagh Quantification of microglial phagocytosis by a flow cytometer-based assay -- Refik Pul, Kandiyil Prajeeth Chittappen, and Martin Stangel Quantification of microglial proliferation and apoptosis by flow cytometry -- Alicia A. Babcock, Martin Wirenfeldt, and Bente Finsen Fluorescence imaging of intracellular Ca2+, Na+, and H+ in cultured microglia -- Tom Schilling and Claudia Eder Patch clamp protocols to study ion channel activity in microglia -- Tom Schilling and Claudia Eder -- Part VI Analysis of microglia polarization -- Studying M1 and M2 states in adult microglia -- Sadanand M. Gaikwad and Michael T. Heneka Isolating, culturing, and polarizing primary human adult and fetal microglia -- Bryce A. Durafourt ... [et al.] -- Part VII Co-culture systems to analysis microglia interactions with other cell types -- Understanding microglia-neuron cross talk : relevance of the microglia-neuron cocultures -- Fernando G. Correa, Miriam Hernangómez, and Carmen Guaza Preparation of rodent primary cultures for neuron-glia, mixed glia, enriched microglia, and reconstituted cultures with microglia -- Shih-Heng Chen, Esteban A. Oyarzabal, and Jau-Shyong Hong -- Part VIII Analysis of microglia functions in vivo -- Microglia detection by enzymatic histochemistry -- Beatriz Almolda, Berta González, and Bernardo Castellano Tomato lectin histochemistry for microglial visualization -- Nàdia Villacampa ... [et al.] Immunohistochemical detection of microglia -- Rocío M. de Pablos, Ana M. Espinosa-Oliva, and Antonio J. Herrera Intrathecal Infusion of Microglia Cells -- Takahiro Masuda ... [et al.] Intracranial injection of LPS in rat as animal model of neuroinflammation -- Ana M. Espinosa-Oliva, Rocío M. de Pablos, and Antonio J. Herrera Analyses of microglia effector function using CX3CR1-GFP knock-in mice -- Jenny A. Garcia, Sandra M. Cardona, and Astrid E. Cardona In vivo two-photon microscopy of microglia -- Satoru Kondo and Shigeo Okabe Use of confocal microscopy in the study of microglia in a brain metastasis model -- Manuel Sarmiento. Front Matter....Pages i-xii Front Matter....Pages 1-1 Front Matter....Pages 3-8 Front Matter....Pages 9-9 Front Matter....Pages 11-16 Front Matter....Pages 17-23 Front Matter....Pages 25-31 Front Matter....Pages 33-39 Front Matter....Pages 41-51 Front Matter....Pages 53-53 Back Matter....Pages 55-61 ....Pages 63-67