A fast and effective microfluidic spraying-plunging method for high-resolution single-particle Cryo-EM

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

A fast and effective microfluidic spraying-plunging method for high-resolution single-particle Cryo-EM

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

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

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.

Original language	English
Title of host publication	Novel Developments in Cryo-EM of Biological Molecules
Subtitle of host publication	Resolution in Time and State Space
Publisher	Taylor and Francis
Pages	395-421
Number of pages	27
ISBN (Electronic)	9781000989441
ISBN (Print)	9789814968768
State	Published - Oct 6 2023

Cite this

Feng, X., Fu, Z., Kaledhonkar, S., Jia, Y., Shah, B., Jin, A., Liu, Z., Sun, M., Chen, B., Grassucci, R. A., Ren, Y., Jiang, H., Frank, J., & Lin, Q. (2023). A fast and effective microfluidic spraying-plunging method for high-resolution single-particle Cryo-EM. In Novel Developments in Cryo-EM of Biological Molecules: Resolution in Time and State Space (pp. 395-421). Taylor and Francis.

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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.",

author = "Xiangsong Feng and Ziao Fu and Sandip Kaledhonkar and Yuan Jia and Binita Shah and Amy Jin and Zheng Liu and Ming Sun and Bo Chen and Grassucci, {Robert A.} and Yukun Ren and Hongyuan Jiang and Joachim Frank and Qiao Lin",

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A fast and effective microfluidic spraying-plunging method for high-resolution single-particle Cryo-EM. / Feng, Xiangsong; Fu, Ziao; Kaledhonkar, Sandip et al.
Novel Developments in Cryo-EM of Biological Molecules: Resolution in Time and State Space. Taylor and Francis, 2023. p. 395-421.

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

TY - CHAP

T1 - A fast and effective microfluidic spraying-plunging method for high-resolution single-particle Cryo-EM

AU - Feng, Xiangsong

AU - Fu, Ziao

AU - Kaledhonkar, Sandip

AU - Jia, Yuan

AU - Shah, Binita

AU - Jin, Amy

AU - Liu, Zheng

AU - Sun, Ming

AU - Chen, Bo

AU - Grassucci, Robert A.

AU - Ren, Yukun

AU - Jiang, Hongyuan

AU - Frank, Joachim

AU - Lin, Qiao

PY - 2023/10/6

Y1 - 2023/10/6

N2 - 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.

AB - 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.

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M3 - Chapter

AN - SCOPUS:85169763342

SN - 9789814968768

SP - 395

EP - 421

BT - Novel Developments in Cryo-EM of Biological Molecules

PB - Taylor and Francis

ER -

A fast and effective microfluidic spraying-plunging method for high-resolution single-particle Cryo-EM

Abstract

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