Single Particle Analysis
Single-particle analysis is the extraction and averaging of individual protein and protein complexes' particles to resolve a 3D structure of the particle in question at near-atomic resolution
Sryo-EM single particle analysis workflow
Purified protein
sample
Preparation of cryo grids
Imaging and data collection
Initial data analysis, 2D, 3D classifications
3D structure reconstruction
Purified protein samples can be provided by the customer or generated by us. We offer protein overexpression and purification services for cryoEM analysis. Contact us to discuss your project!
The purified protein is applied to a cryo-grid and flash-frozen in liquid ethane. At high magnification on a cryo-TEM, a single day of automated data collection produces thousands of images, each with hundreds of particles.
Our data analysis software allows for the extraction and filtering of the particles in each image, ensuring that only the best particle images are incorporated into the final reconstruction.
Particle filtering is initially done through 2D classification. The secondary structure of proteins is often already visible at this step.
After selecting particles optimal for 3D averaging, a low-resolution volume is generated from a subset of the data. This volume will be used to align each particle for high-resolution reconstruction
After the generation of a low-resolution volume, further refinement with all optimal particles is performed to resolve a high-resolution structure. Our method allows achieving structures with sub 2 Å resolution.