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![[Figure 1]](ai5014fig1mag.jpg)
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Figure 1
Overview of the cryo-EM workflow. Step 1: evaluation of protein sample purity and quality to determine suitability for structural characterization by cryo-EM. Step 2: vitrification of samples for cryo-EM imaging by applying the purified protein to a cryo-EM grid followed by plunge-freezing. Step 3: screening of vitrified cryo-EM grids to identify optimal freezing conditions for the purified sample. Parameters such as grid type, buffer components, additives and plunge-freezing time/force are assessed. Once sample preparation is optimized, larger cryo-EM data sets are collected. Step 4: data processing using specialized software, including motion correction, CTF estimation, particle picking, 2D classification and 3D refinement, to obtain a high-resolution 3D map. Step 5: model building and refinement to fit a model into the map. Model validation is performed to assess the quality of the map and the model fit. Note: the cryo-EM workflow often requires iterative optimization at multiple stages, and users may need to revisit earlier steps to achieve a final structure at the desired resolution. |
![[Figure 1]](ai5014fig1.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
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