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Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition
This content is Free Access.
Chapters
- 00:05Title
- 01:16Making Carbon Coated EM Grids for Negative Staining EM
- 02:28Carbon Coating Grids
- 04:27Preparing Negative Stain EM Grids
- 05:57Preparing an Electron Microscope
- 07:07Electron Microscopy Images
- 07:30Conclusion
Visualizing protein samples by negative stain electron microscopy (EM) has become a popular structural analysis method. It is useful for quantitative structural analysis, such as calculating a 3D reconstruction of the molecules being studied, and also for qualitative examination of the quality of protein preparations. In this article we present detailed protocols for preparing the EM grids, staining the sample and visualizing the sample in an electron microscope. Novice users can follow these protocols easily and to utilize negative stain EM as a routine assay, in addition to other biochemical assays, for evaluating their protein samples.
Tags
VisualizingProteinsMacromolecular ComplexesNegative Stain EMGrid PreparationImage AcquisitionSingle Particle Electron MicroscopyStructural BiologyProtein StructuresCryoEMSample Preparation MethodDried Heavy Metal SaltSpecimen ContrastNegative Stain EMThree-dimensional StructurePurified ProteinsProtein ComplexesHomogeneity/heterogeneityLarge AssembliesQuality EvaluationEM ProtocolCarbon Coated GridsElectron Microscope
