High-Resolution Imaging of Cellular Proteins

• Expression of Epitope-Tagged Proteins in Mammalian Cells in Culture.- Antibody Production with Synthetic Peptides.- Production and Purification of Polyclonal Antibodies.- Preparation of Colloidal Gold Particles and Conjugation to Protein A/G/L, IgG, F(ab’)2 and Streptavidin.- Helper Dependent Adenoviral Vectors and their use for Neuroscience Applications.- Localizing Proteins in Fixed Giardia Lamblia and Live Cultured Mammalian Cells by Confocal Fluorescence Microscopy.- Using Fluorescent Protein Fusions to Study Protein Subcellular Localization and Dynamics in Plant Cells.- Using FRAP Or FRAPA to Visualize the Movement of Fluorescently-Labeled Proteins or Cellular Organelles in Live Cultured Neurons Transformed with Adeno-Associated Viruses.- Bimolecular Fluorescence Complementation (BiFc) Analysis of Protein-Protein Interactions and Assessment of Subcellular Localization in Live Cells.- Viral Injection and Cranial Window Implantation for In Vivo Two-Photon Imaging.- Imaging SynapticVesicle Exocytosis-Endocytosis with pH Sensitive Fluorescent Proteins.- Immunogold Protein Localization on Grid-Glued Freeze-Fracture Replicas.- Pre-Embedding Double-Label Immunoelectron Microscopy of Chemically Fixed Tissue Culture Cells.- Immunoelectron Microscopy of Cryofixed and Freeze- Substituted Plant Tissues.- Immunoelectron Microscopy of Cryofixed Freeze Substituted Yeast Saccharomyces cerevisiae.- Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes.- Pre-Embedding Nanogold Silver and Gold Intensification.- Post-Embedding Mammalian Tissue for Immunoelectron Microscopy: A Standardized Procedure Based On Heat-Induced Antigen Retrieval.- Pre- and Post-Embedding Immunogold Labeling of Tissue Sections.- Immuno-Gold Labelling for Scanning Electron Microscopy.- Monitoring Synaptic Vesicle Protein Sorting with Enhanced Horseradish Peroxidase in the Electron Microscope.- Functional Nanoscale Imaging Of Synaptic VesicleCycling With Superfast Fixation.

“This is a collection of timely techniques and methods that have been key for the development of imaging at the structural and ultrastructural level. … The intended audience is cell biologists and all scientists working with imaging techniques to localize protein expression at a cellular and subcellular level. … Each chapter ends with a list of references. Graphics and high-quality figures are displayed where needed.” (Michele Fornaro, Doody's Book Reviews, July, 2017)