Immunogold Labeling for Scanning Electron Microscopy
- 12 August 2016
- book chapter
- protocol
- Published by Springer Science and Business Media LLC in Methods in molecular biology (Clifton, N.J.)
- Vol. 1474, 309-325
- https://doi.org/10.1007/978-1-4939-6352-2_20
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.This publication has 9 references indexed in Scilit:
- Imaging Yeast NPCsMethods in Cell Biology, 2014
- Entry into the nuclear pore complex is controlled by a cytoplasmic exclusion zone containing dynamic GLFG-repeat nucleoporin domainsJournal of Cell Science, 2013
- Filaments made from A- and B-type lamins differ in structure and organizationJournal of Cell Science, 2008
- Visualization of the nucleus and nuclear envelope in situ by SEM in tissue culture cellsNature Protocols, 2007
- Electron Microscopic Analysis of the Leading Edge in Migrating CellsMethods in Cell Biology, 2007
- Yeast nuclear pore complexes have a cytoplasmic ring and internal filamentsJournal of Structural Biology, 2004
- High‐resolution field‐emission scanning electron microscopy of nuclear pore complexScanning, 1997
- Three-Dimensional Visualization of the Route of Protein Import: The Role of Nuclear Pore Complex SubstructuresExperimental Cell Research, 1997
- High resolution scanning electron microscopy of the nuclear envelope: demonstration of a new, regular, fibrous lattice attached to the baskets of the nucleoplasmic face of the nuclear pores.The Journal of cell biology, 1992