Category Ranking
98%
Total Visits
921
Avg Visit Duration
2 minutes
Citations
20
Article Abstract
We present an approach for preparing cryo-electron microscopy (cryo-EM) grids to study short-lived molecular states. Using piezoelectric dispensing, two independent streams of ~50-pl droplets of sample are deposited within 10 ms of each other onto the surface of a nanowire EM grid, and the mixing reaction stops when the grid is vitrified in liquid ethane ~100 ms later. We demonstrate this approach for four biological systems where short-lived states are of high interest.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7799389 | PMC |
| http://dx.doi.org/10.1038/s41592-020-0925-6 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Amyloid-β oligomers, curvilinear and annular assemblies, imaged by cryo-ET, cryo-EM, and AFM.
Sci Adv
August 2025
Department of Biochemistry, School of Biological and Behavioural Sciences, Queen Mary University of London, London, UK.
Prefibrillar structures of the amyloid-β (Aβ) peptide are central to cytotoxicity in Alzheimer's disease. Time-resolved imaging of oligomers has enabled quantification of their extension. A snapshot of these prefibrillar assemblies has been characterized using a combination of cryo-electron tomography (cryo-ET), cryo-electron microscopy (cryo-EM) single-particle analysis, and atomic force microscopy (AFM).
View Article and Find Full Text PDFLaser flash melting cryo-EM samples to overcome preferred orientation.
Nat Methods
August 2025
Ecole Polytechnique Fédérale de Lausanne (EPFL), Laboratory of Molecular Nanodynamics, Lausanne, Switzerland.
Sample preparation remains a bottleneck for protein structure determination by cryo-electron microscopy. A frequently encountered issue is that proteins adsorb to the air-water interface of the sample in a limited number of orientations. This makes it challenging to obtain high-resolution reconstructions, or may even cause projects to fail altogether.
View Article and Find Full Text PDFTime-Resolved Fluorescence Imaging and Correlative Cryo-Electron Tomography to Study Structural Changes of the HIV-1 Capsid.
ACS Nano
September 2025
Institute of Molecular Biophysics and Department of Biological Sciences, Florida State University, Tallahassee, Florida 32306, United States.
The conical HIV-1 capsid protects the internal viral genome and facilitates the infection of target cells. Highly potent antivirals, such as the clinically approved drug Lenacapavir (LEN), block HIV-1 replication by changing the capsid structure and modulating its function. However, structural studies of the HIV-1 capsid, its disassembly, or stabilization by antivirals have been challenging.
View Article and Find Full Text PDFMix-it-up: Accessible time-resolved cryo-EM on the millisecond timescale.
bioRxiv
July 2025
Department of Integrative Structural and Computational Biology, Scripps Research; La Jolla, CA 92037, USA.
Biological reactions often involve macromolecules that undergo substrate-induced conformational changes in under a second, yet capturing these transient states remains challenging. While high-resolution structural techniques such as X-ray crystallography and cryo-electron microscopy (cryo-EM) have advanced our mechanistic understanding of protein-substrate interactions, traditional sample preparation methods are too slow to capture rapid biochemical events. Time-resolved cryo-EM has emerged as a promising approach to visualize structural dynamics on microsecond-to-millisecond timescales, but its widespread adoption has been limited by costly equipment and challenges in achieving rapid mixing, application, and vitrification of samples in a reproducible manner.
View Article and Find Full Text PDFUltrathin Liquid Cells for Microsecond Time-Resolved Cryo-EM.
bioRxiv
May 2025
Ecole Polytechnique Fédérale de Lausanne (EPFL), Laboratory of Molecular Nanodynamics, CH-1015 Lausanne, Switzerland.
Microsecond time-resolved cryo-electron microscopy promises to significantly advance our understanding of protein function by rendering cryo-electron microscopy (cryo-EM) fast enough to observe proteins at work. This emerging technique involves flash melting a cryo sample with a laser beam to provide a brief time window during which dynamics are initiated. When the laser is switched off, the sample revitrifies, arresting the proteins in their transient configurations.
View Article and Find Full Text PDF