scholarly journals Self-Blotting Nanowire Grids for Cryo-EM Sample Preparation

2017 ◽  
Vol 23 (S1) ◽  
pp. 848-849
Author(s):  
Hui Wei ◽  
Venkat Dandey ◽  
Zhening Zhang ◽  
Ashleigh Raczkowski ◽  
Bridget Carragher ◽  
...  
Keyword(s):  
Thin Film ◽  
Electron Microscopy ◽  
Sample Preparation ◽  
Filter Paper ◽  
Ice Thickness ◽  
New Approach ◽  
Cryo Electron Microscopy ◽  
Piezo Electric

AbstractAlmost every aspect of cryo electron microscopy (CryoEM) has been automated over the last few decades. One of the challenges that remains to be addressed is the robust and reliable preparation of vitrified specimens of suitable ice thickness. The development of a new self-blotting nanowire (Zhang et al., 2013) grid in conjunction with a piezo electric dispensing robot called Spotiton (Jain et al., 2012) enables spreading a sample to a thin film without the use of externally applied filter paper. This new approach has the advantage of using small amounts of protein material, resulting in large areas of ice of a well- defined thickness containing evenly distributed particles (Razinkov et al., 2016).

Tailoring surface acoustic wave atomisation for cryo-electron microscopy sample preparation

Lab on a Chip ◽  
2019 ◽  
Vol 19 (8) ◽  
pp. 1378-1385 ◽  
Author(s):  
Dariush Ashtiani ◽  
Alex de Marco ◽  
Adrian Neild
Keyword(s):  
Electron Microscopy ◽  
Fluid Flow ◽  
Sample Preparation ◽  
Acoustic Wave ◽  
Surface Acoustic Wave ◽  
Flow Rate ◽  
Fluid Flow Rate ◽  
Cryo Electron Microscopy ◽  
Electron Microscopy Grid

Surface acoustic wave (SAW) atomisation is investigated in the context of cryo electron microscopy grid preparation. Here, the primary requirements are a reproducible and narrow plume of droplets delivering a low fluid flow rate.

Cryogenic Transmission Electron Microscopy: Aqueous Suspensions of Nanoscale Objects

2013 ◽  
Vol 19 (6) ◽  
pp. 1542-1553 ◽  
Author(s):  
Nathan D. Burrows ◽  
R. Lee Penn
Keyword(s):  
Thin Film ◽  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Sample Preparation ◽  
Self Assembly ◽  
Aqueous Samples ◽  
Liquid Media ◽  
Cryogenic Transmission Electron Microscopy ◽  
Transmission Electron ◽  
The Many

AbstractDirect imaging of nanoscale objects suspended in liquid media can be accomplished using cryogenic transmission electron microscopy (cryo-TEM). Cryo-TEM has been used with particular success in microbiology and other biological fields. Samples are prepared by plunging a thin film of sample into an appropriate cryogen, which essentially produces a snapshot of the suspended objects in their liquid medium. With successful sample preparation, cryo-TEM images can facilitate elucidation of aggregation and self-assembly, as well as provide detailed information about cells and viruses. This work provides an explanation of sample preparation, detailed examples of the many artifacts found in cryo-TEM of aqueous samples, and other key considerations for successful cryo-TEM imaging.

scholarly journals Antibody-Based Affinity Cryo-Electron Microscopy at 2.6 Å Resolution

2016 ◽  
Author(s):  
Guimei Yu ◽  
Kunpeng Li ◽  
Pengwei Huang ◽  
Xi Jiang ◽  
Wen Jiang
Keyword(s):  
Electron Microscopy ◽  
Sample Preparation ◽  
High Concentration ◽  
Low Concentration ◽  
Atomic Structures ◽  
Cryo Electron Microscopy ◽  
Grid Approach ◽  
Reconstruction Quality ◽  
Tulane Virus ◽  
Quantitative Analyses

AbstractThe affinity cryo-electron microscopy (cryo-EM) approach has been explored in recent years to simplify and improve the sample preparation for cryo-EM. Despite the demonstrated successes for low-concentration and unpurified specimens, the lack of near-atomic structures using this approach has led to a common perception of affinity cryo-EM as a niche technique incapable of reaching high resolutions. Here, we report a ~2.6 Å structure solved using the antibody-based affinity grid approach with a Tulane virus sample of low concentration. This is the first near-atomic structure solved using the affinity cryo-EM approach. Quantitative analyses of the structure indicate data and reconstruction quality comparable to conventional grid preparation method using samples at high concentration. With the shifting of bottlenecks of cryo-EM structural studies to sample grid preparation, our demonstration of the sub-3 Å capability of affinity cryo-EM approach indicates its potential in revolutionizing cryo-EM sample preparation for a broader spectrum of specimens.

scholarly journals An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology

Methods ◽  
2016 ◽  
Vol 100 ◽  
pp. 3-15 ◽  
Author(s):  
Rebecca F. Thompson ◽  
Matt Walker ◽  
C. Alistair Siebert ◽  
Stephen P. Muench ◽  
Neil A. Ranson
Keyword(s):  
Electron Microscopy ◽  
Sample Preparation ◽  
Structural Biology ◽  
Cryo Electron Microscopy

scholarly journals Amorphous nickel titanium alloy film: a new choice for cryo electron microscopy sample preparation

2020 ◽  
Author(s):  
Xiaojun Huang ◽  
Lei Zhang ◽  
Zuoling Wen ◽  
Hui Chen ◽  
Shuoguo Li ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Titanium Alloy ◽  
Data Collection ◽  
Sample Preparation ◽  
Particle Distribution ◽  
Carbon Film ◽  
Nickel Titanium ◽  
Pure Gold ◽  
Interface Problem ◽  
Cryo Electron Microscopy

ABSTRACTCryo-electron microscopy (cryoEM) has become one of the most important approach for structural biology. However, barriers are still there for an increase successful rate, a better resolution and improved efficiency from sample preparation, data collection to image processing. CryoEM sample preparation is one of the bottlenecks with many efforts made recently, including the optimization of supporting substrate (e.g. ultra-thin carbon, graphene, pure gold, 2d crystal of streptavidin, and affinity modification), which was aimed to solve air-water interface problem, or reduce beam induced motion (BIM), or change particle distribution in the grid hole. Here, we report another effort of developing a new supporting substrate, the amorphous nickel-titanium alloy (ANTA) film, for cryoEM sample preparation. Our investigations showed advantages of ANTA film in comparison with conventional carbon film, including superior electron conductivity and trace non-specific interaction with protein. These advantages yield less BIM and significantly improved particle distribution during cryoEM experiment of human apo-ferritn, thus resulting an improved reconstruction resolution from a reduced number of micrographs and particles. Unlike the pure gold film, the usage of the ANTA film is just same with the carbon film, compatible to conventional automatic cryoEM data collection procedure.

scholarly journals An open-source cryo-storage solution

2017 ◽  
Author(s):  
Eveline Ultee ◽  
Fred Schenkel ◽  
Wen Yang ◽  
Susanne Brenzinger ◽  
Jamie S. Depelteau ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Sample Preparation ◽  
Structural Biology ◽  
Storage Capacity ◽  
Storage System ◽  
Cost Effective ◽  
Wait Times ◽  
Sample Storage ◽  
Cryo Electron Microscopy ◽  
And Storage

AbstractThe field of cryo-electron microscopy is a rapidly growing method in structural biology. With this development, access to cryo-EM facilities becomes a bottleneck that results in long wait times between sample preparation and data acquisition. To improve sample storage, we developed a cryo-storage system with a more efficient and larger storage capacity that enables cryo-sample storage in a highly organized manner. This system is simple to use, cost-effective and easily adaptable for any type of grid box and storage dewar and any size cryo-EM laboratory.

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