Guide to Research Techniques in Neuroscience

Name: Guide to Research Techniques in Neuroscience
Price: 5.7 USD
Availability: InStock
ISBN: 9780123748492

ISBN-10: 0123748496

ISBN-13: 9780123748492

Edition: 2010

Authors: Matt Carter, Jennifer C. Shieh

List price: $65.95

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Description:

Neuroscience is by definition a multidisciplinary field: some scientists study genes and proteins at the molecular level while others study neural circuitry. A single topic such as the auditory system can be studied using techniques from genetics, imaging, biochemistry, or electrophysiology. A young scientist must learn how to read the primary literature and then develop their own experiments. This book offers that scientist an overview of mainstream research techniques, provides guidelines on how to choose one technique over another, offers tips on analyzing data, and provides a list of references for additional detailed study. This book can also assist an experienced scientist understand…

Book details

List price: $65.95
Copyright year: 2010
Publisher: Elsevier Science & Technology
Publication date: 10/22/2009
Binding: Paperback
Pages: 408
Size: 5.94" wide x 9.00" long x 0.75" tall
Weight: 1.430
Language: English

Matt Carter, PhD, is currently Assistant Professor of Biology at Williams College. His previous position was as a post-doctoral fellow in Richard Palmiter's lab at the University of Washington using optogenetic techniques to study neural circuitry. He has authored the first edition of this book (Elsevier, 2009) as well as Designing Science Presentations: A Visual Guide to Figures, Papers, Slides, Posters, and More (Elsevier, 2012). He was the awardee of Stanford University's Walter J. Gores Award for Excellence in Teaching, and two-time recipient of the Stanford School of Medicine's Excellence in Teaching Award. He currently teaches courses at Williams in both Topics in Neuroscience as well…



Preface


Foreword


Introduction



Whole Brain Imaging


Structural Brain Imaging Techniques


Cerebral Angiography


Computerized Tomography (CT)


Magnetic Resonance Imaging (MRI)


Diffusion Magnetic Resonance imaging (diffusion MRI)


Functional Brain Imaging Techniques


Functional Magnetic Resonance Imaging (FMRI)


Positron Emission Tomography (PET)


Single-Proton Emission Computerized Tomography (SPECT)


Electroencephalography (EEC)


Magnetoencephalography (MEG)


Optical Imaging


Functional Imaging Experimental Design and Analysis


Planning the Experiment


Conducting the Experiment


Manipulating Neural Activity during an Experiment


Postexperimental Data Analysis


Conclusion


Suggested Reading and References



Animal Behavior


Considerations for Choosing and Performing a Behavioral Assay


Choosing an Appropriate Model Organism


Choosing an Appropriate Behavioral Paradigm


Variability in Individuals


Using Animal Behavior as a Model for Human Behavior


Rodent Behavioral Paradigms


Locomotor Activity


Motor Coordination and Balance


Sensory Function


Nociception


Spatial Learning and Memory


Nonspatial Learning and Memory


Social Behaviors


Anxiety


Depression


Drosophila Behavioral Paradigms


Locomotor Behavior


Flight


Sensory Function


Learning and Memory


Social Behaviors


C. elegans Behavioral Paradigms


Locomotor Behavior


Sensory Behavior


Nonhuman Primate Behavioral Paradigms


Conclusion


Suggested Reading and References



Stereotaxic Surgeries and In Vivo Techniques


Stereotaxic Surgeries


Stereotaxic Surgeries in Rodents


Stereotaxic Surgeries in Nonhuman Primates


Implants for Long-Term Access to the Brain


Sealable Chambers


Cannulae


Cranial Windows


Measuring Neural Activity In Vivo


Electrophysiology


Fluorescent Activity Indicators


Measuring Neurochemistry in Vivo


Microdialysis


Voltammetry and Amperometry


Manipulating the Brain In Vivo


Physical Manipulation


Pharmacological Manipulation


Electrical Manipulation


Genetic Manipulation


Conclusion


Suggested Reading and References



Electrophysiology


A Brief Review of the Electrical Properties of Neurons


The Electrophysiology Rig


Types of Electrophysiology Recordings


Extracellular Recording


Intracellular Recording


Patch Clamp Techniques


Electrophysiology Tissue Preparations


In Vitro Recordings


In Vivo Recordings


Methods of Manipulating Neurons During Electrophysiology Experiments


Conclusion


Suggested Reading and References



Microscopy


Essential Principles of Microscopy


Fundamental Parameters in Microscopy


The Design of a Compound Microscope


The Design of a Stereomicroscope


Light Microscopy


Fluorescence Microscopy


Epifluorescent Microscopy


Confocal Microscopy


Two-Photon Microscopy


Total Internal Reflection Fluorescence (TIRF) Microscopy


Electron Microscopy


Transmission Electron Microscopy (TEM)


Scanning Electron Microscopy (SEM)


Electron Tomography (ET)


Preparing and Interpreting Microscopy Data


Image Processing


Interpreting Images


Conclusion


Suggested Reading and References



Visualizing Nervous System Structure


Tissue Preparation


Fixation


Embedding


Sectioning


Visualizing Morphology


Cell Body Stains


Fiber Stains


Golgi Stain


Intracellular and Juxtacellular Labeling


Fluorescent Labels


Chromogenic/Colorimetric Labels


Radioactive Labels


Cold Labels


Visualizing Gene and Protein Expression


In Situ Hybridization


Immunohistochemistry


Enzymatic Histochemistry


Reporter Genes


Visualizing Circuitry


Anterograde and Retrograde Tracers


Transsynaptic Labeling


Conclusion


Suggested Reading and References



Visualizing Nervous System Function


Static Markers of Activity


Assaying Neural Activity in Fixed Tissue


Assaying Cellular Function in Fixed Tissue


Visualizing Neural Activity


Imaging Voltage


Imaging Calcium


Imaging Synaptic Transmission


Optically Manipulating Neural Activity


Stimulation through Uncaging of Molecules


Light-Activated Channels


Visualizing Protein Function


Time-Lapse Imaging with Reporter Genes


Fluorescence Resonance Energy Transfer (FRET)


Bimolecular Fluorescence Complementation (BiFC)


Fluorescence Recovery after Photo bleaching (FRAP)


Photoactivation and Photoconversion


Optically Manipulating Protein Function


Photoactivation/Photo-uncaging


Photoinactivation


Conclusion


Suggested Reading and References



Identifying Genes and Proteins of Interest


A Brief Review of Genes and Proteins


The Central Dogma of Molecular Biology


DNA


Transcription


Translation


Genetic Screens


Forward Genetic Screen


Reverse Genetic Screen


In Silico Screens


Blast


Ensembl


Molecular Screens


cDNA Microarray Screen


RNAi Screen


Conclusion


Suggested Reading and References



Molecular Cloning and Recombinant DNA Technology


Isolating DNA Fragments


Restriction Enzymes


Polymerase Chain Reaction (PCR)


Cloning DNA


Vectors


Ligation


Transformation


Purifying DNA


Isolation and Characterization of DNA Fragments Using Gel Electrophoresis


Purifying DNA from Host Cells


Identifying DNA


DNA Sequencing


Nucleic Acid Hybridization Techniques


Conclusion


Suggested Reading and References



Gene Delivery Strategies


Physical Gene Delivery


Microinjection


Electroporation


Biolistics


Chemical Gene Delivery


Calcium Phosphate Transfection


Lipid Transfection


Viral Gene Delivery


Adenovirus


Adeno-Associated Virus (AAV)


Lentivirus


Herpes Simplex Virus (HSV)


Conclusion


Suggested Reading and References



Making and Using Transgenic Organisms


Transgenes


Reporter Genes


Genes Used to Ablate Neurons


Genes Used to Measure Neural Activity


Genes Used to Manipulate Neural Activity


Genes Used to Disrupt Endogenous Gene Function


Overexpression or Misexpression of Endogenous Genes


The Transgenic Construct


Binary Transgenic Systems


The Gal4/UAS System


The Cre/Lox System


The Flp/Frt System


TheTet-off/Tet-on System


Making Transgenic Organisms


Making Transgenic Mice


Making Transgenic Flies


Making Transgenic Worms


Making Other Transgenic Organisms


Conclusion


Suggested Reading and References



Manipulating Endogenous Genes


Genetic Targeting


Knockout


Knockin


Conditional Knockout


Disrupting Gene Products


RNA Interference (RNAi)


Morpholtnos


Dominant Negatives


Conclusion


Suggested Reading and References



Cell Culture Techniques


Cell Culture Equipment and Reagents


Equipment


Culture Media


Immortalized Cell Lines


Primary Cell and Tissue Culture


Dissociated Cell Cultures


Slice Cultures


Explant Cultures


Stem Cell Cultures


Embryonic Stem Cells


Neural Stem Cells


Induced Plurrpotent Stem Cells


Manipulating Cells in Culture


Transfection


Coculture Systems


Pharmacology


Antibody Interference


Conclusion


Suggested Reading and References



Biochemical Assays and Intracellular Signaling


Introduction to Signal Transduction and Intracellular Signaling


Fundamental Tools for Studying Proteins


Making and Using Antibodies


Purifying Proteins


Immunoprecipitation (IP)


Investigating Protein Expression


Western Blot (WB)


Enzyme-Linked Immunosorbent Assay (ELISA)


Radioimmunoassay (RIA)


Immunohistochemistry (IHC)


Immunoelectron Microscopy (IEM)


Reporter Proteins


Investigating Protein-Protein Interactions


Coimmunoprecipitation (Co-IP)


Protein Affinity Chromatography


Yeast Two-Hybrid Assay


Investigating Posttranslational Modifications


PTM-Specific Assays


PTM-Specific Antibodies


Investigating Protein-DNA Interactions


Electrophoretic Mobility Shift Assay (EMSA)


Chromatin Immunoprecipitation (ChIp)


Luciferase Assay


Conclusion


Suggested Reading and References


Glossary


Index