title: Immunogold Labeling for Scanning Electron Microscopy.
abstract: Scanning electron microscopes are useful biological tools that can be used to image the surface of whole organisms, tissues, cells, cellular components, and macromolecules. Processes and structures that exist at surfaces can be imaged in pseudo, or real 3D at magnifications ranging from about 10× to 1,000,000×. Therefore a whole multicellular organism, such as a fly, or a single protein embedded in one of its cell membranes can be visualized. In order to identify that protein at high resolution, or to see and quantify its distribution at lower magnifications, samples can be labeled with antibodies. Any surface that can be exposed can potentially be studied in this way. Presented here is a generic method for immunogold labeling for scanning electron microscopy, using two examples of specimens: isolated nuclear envelopes and the cytoskeleton of mammalian culture cells. Various parameters for sample preparation, fixation, immunogold labeling, drying, metal coating, and imaging are discussed so that the best immunogold scanning electron microscopy results can be obtained from different types of specimens.
authors: School of Biological and Biomedical Sciences, Durham University, South Road, Durham, DH1 3LE, UK. m.w.goldberg@durham.ac.uk.
authors: Martin W
authors: MW
authors: Goldberg
authors: Department of Biology of the Cell Nucleus, Institute of Molecular Genetics AS CR, v.v.i.,, Videnska 1083, 142 20, Prague 4, Czech Republic.
authors: Jindřiška
authors: J
authors: Fišerová
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D000941:Antigens / Q000235:genetics* / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003599:Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure; D004853:Epoxy Resins / Q000737:chemistry; D005260:Female; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D015870:Gene Expression; D017979:Gold Colloid / Q000737:chemistry*; D007150:Immunohistochemistry / Q000379:methods*; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D008867:Microtomy; D009685:Nuclear Envelope / Q000378:metabolism / Q000648:ultrastructure; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism / Q000648:ultrastructure; D011108:Polymers / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D016610:Tissue Embedding / Q000379:methods; D016707:Tissue Fixation / Q000379:methods*; D014982:Xenopus laevis
chemical_list: D000906:Antibodies; D000941:Antigens; D004853:Epoxy Resins; D005404:Fixatives; D017979:Gold Colloid; D028861:Nuclear Pore Complex Proteins; D011108:Polymers; D005557:Formaldehyde; C003043:paraform
keywords: Envelope; Immunogold; Labeling; Nuclear; Scanning electron microscopy; Xenopus