Structural details of individual particles can be difficult to discern from raw images taken on an electron microscope. This is due to the low signal-to-noise ratio typical for images of radiation-sensitive biological samples. It becomes even more challenging when the particles are conformationally or compositionally heterogeneous.
2D class averaging is the first step in visualizing the three dimensional structure of a protein or macromolecule, and can provide researchers a comprehensive representation of particles in their sample. 2D class averages can help visually assess variability in qualities such as preferred orientation, conformation, aggregation, and homogeneity of a sample in order to help select the most viable samples for full 3D structure reconstruction.
Types of Proteins that can be 2D Class Averaged
Frequently Asked Questions
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Can I see an example report showing Negative Stain or Cryo-EM 2D class averages?
Yes! Contact us to see an example of our Starter Pack, or feasibility assessment, performed on B-amylase, which includes negative stain screening, 2D classifications in negative stain and cryo-EM, and an initial 3D structure reconstruction.
When do you use negative stain imaging versus cryo-EM imaging for obtaining 2D classifications?
When deciding whether to use negative stain imaging or cryo-EM imaging to obtain 2D class averages, our scientists take multiple elements into consideration, including the sample type, sample stability, and sample concentration.
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