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Detection of Microcrystals for CryoEM.
Weiss, Simon; Vergara, Sandra; Lin, Guowu; Calero, Guillermo.
Affiliation
  • Weiss S; Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
  • Vergara S; Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
  • Lin G; Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
  • Calero G; Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. guc9@pitt.edu.
Methods Mol Biol ; 2215: 299-307, 2021.
Article in En | MEDLINE | ID: mdl-33368010
Here, we present a strategy to identify microcrystals from initial protein crystallization screen experiments and to optimize diffraction quality of those crystals using negative stain transmission electron microscopy (TEM) as a guiding technique. The use of negative stain TEM allows visualization along the process and thus enables optimization of crystal diffraction by monitoring the lattice quality of crystallization conditions. Nanocrystals bearing perfect lattices are seeded and can be used for MicroED as well as growing larger crystals for X-ray and free electron laser (FEL) data collection.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Protein Conformation / Cryoelectron Microscopy / Microscopy, Electron, Transmission / Nanoparticles Type of study: Diagnostic_studies Language: En Journal: Methods Mol Biol Journal subject: BIOLOGIA MOLECULAR Year: 2021 Type: Article Affiliation country: United States

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Protein Conformation / Cryoelectron Microscopy / Microscopy, Electron, Transmission / Nanoparticles Type of study: Diagnostic_studies Language: En Journal: Methods Mol Biol Journal subject: BIOLOGIA MOLECULAR Year: 2021 Type: Article Affiliation country: United States