A Fast and Effective Microfluidic Spraying-Plunging Method for High-Resolution Single-Particle Cryo-EM

Overview

Journal Structure

Publisher Cell Press

Specialties Biochemistry
Biotechnology
Cell Biology
Molecular Biology

Date 2017 Mar 14

PMID 28286002

Citations 64

Authors

Xiangsong Feng

Ziao Fu

Sandip Kaledhonkar

Yuan Jia

Binita Shah

Amy Jin

Zheng Liu

Ming Sun

Bo Chen

Robert A Grassucci

YuKun Ren

Hongyuan Jiang

Joachim Frank

Qiao Lin

Affiliations

Soon will be listed here.

Abstract

We describe a spraying-plunging method for preparing cryoelectron microscopy (cryo-EM) grids with vitreous ice of controllable, highly consistent thickness using a microfluidic device. The new polydimethylsiloxane (PDMS)-based sprayer was tested with apoferritin. We demonstrate that the structure can be solved to high resolution with this method of sample preparation. Besides replacing the conventional pipetting-blotting-plunging method, one of many potential applications of the new sprayer is in time-resolved cryo-EM, as part of a PDMS-based microfluidic reaction channel to study short-lived intermediates on the timescale of 10-1,000 ms.

Citing Articles

A PDMS-based Microfluidic Chip Assembly for Time-Resolved Cryo-EM (TRCEM) Sample Preparation.

Feng X, Frank J Bio Protoc. 2025; 15(4):e5193.

PMID: 40040794 PMC: 11877145. DOI: 10.21769/BioProtoc.5193.

Time-resolved cryo-EM (TRCEM) sample preparation using a PDMS-based microfluidic chip assembly.

Feng X, Frank J bioRxiv. 2024; .

PMID: 39713383 PMC: 11661195. DOI: 10.1101/2024.12.08.627437.

RNA sample optimization for cryo-EM analysis.

Chen X, Wang L, Xie J, Nowak J, Luo B, Zhang C Nat Protoc. 2024; .

PMID: 39548288 DOI: 10.1038/s41596-024-01072-1.

Preparation of oxygen-sensitive proteins for high-resolution cryoEM structure determination using (an)aerobic blot-free vitrification.

Cook B, Narehood S, McGuire K, Li Y, Tezcan F, Herzik Jr M bioRxiv. 2024; .

PMID: 39091810 PMC: 11291078. DOI: 10.1101/2024.07.19.604374.

Tuning ice thickness using the chameleon for high-quality cryoEM data collection.

McGuire K, Cook B, Narehood S, Herzik Jr M bioRxiv. 2024; .

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