Understanding the invisible hands of sample preparation for cryo-EM
2021年5月7日 · Grid preparation describes the steps needed to make a sample suitable for analysis in the microscope. These steps involve chemical or plasma treatment of the grid, sample deposition and...
Cryo-EM grid optimization for membrane proteins - ScienceDirect
2021年3月19日 · Cryo-EM grid preparation is an important bottleneck in protein structure determination, especially for membrane proteins, typically requiring screening of a large number of conditions. We systematically investigated the effects of buffer components, blotting conditions and grid types on the outcome of grid preparation of five different membrane ...
A strategic approach for efficient cryo-EM grid optimization using ...
2024年2月15日 · The proper optimization of cryo-EM grid quality is essential for ensuring that valuable data collection time is well-utilized and that the best-possible results are obtained, but the process can be time-consuming and resource-exhaustive if …
Functionalized graphene grids with various charges for single
2022年11月7日 · By applying onto three different types of macromolecules, we demonstrate that the high-yield charged graphene grids bring macromolecules away from the AWI and enable adjustable particle-orientation...
Functionalized graphene-oxide grids enable high-resolution cryo-EM ...
2024年3月12日 · Here we present a straightforward and reproducible method for determining structures of the SNF2h-nucleosome complex using graphene-oxide (GO) grids functionalized with negatively charged DNA or...
Batch Production of High-Quality Graphene Grids for Cryo-EM: Cryo-EM …
2023年3月16日 · To demonstrate their practical application, we determined the cryo-EM structure of Methylococcus capsulatus soluble methane monooxygenase hydroxylase (sMMOH) at resolutions of 2.9 and 2.5 Å using Quantifoil and graphene-coated grids, respectively.
Cryo-EM Grid Preparation of Membrane Protein Samples for …
Cryo-EM Grid Preparation. Cryogenic sample-grid preparation allows fixing biological samples by rapidly transferring and cooling them in liquid ethane (−188°C). Under these conditions, ice crystals are unable to form, thus preserving the specimen integrity.
Cryo-EM structure of a photosystem I variant containing an
3 天之前 · Cryo-EM grid preparation and screening. Three microliters of the MenB PS I sample at 2 mg Chl ml –1 was pipetted onto a glow discharged (30 s, 25 mA) Au C-flat 2/1 300-mesh microscopy grid (Electron Microscopy Sciences). A Thermo Fisher Scientific Vitrobot Mark IV system set to 100% humidity and 4°C was used to blot and plunge freeze the ...
Cryo-EM grid optimization for membrane proteins - Cell Press
Cryo-EM grid preparation is an important bottleneck in protein structure determination, especially for membrane proteins, typically requiring screening of a large number of conditions. We systematically investigated the effects of buffer components, blotting conditions and grid types on the outcome of grid preparation of five different membrane ...
High-yield monolayer graphene grids for near-atomic resolution ... - PNAS
2019年12月26日 · Here, we describe a method for preparing graphene cryo-EM grids with up to 99% monolayer graphene coverage that allows for more than 70% grid squares for effective data acquisition with improved image quality and protein density.