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

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",

year = "2023",

month = oct,

day = "6",

language = "English",

isbn = "9789814968768",

pages = "395--421",

booktitle = "Novel Developments in Cryo-EM of Biological Molecules",

publisher = "Taylor and Francis",

}

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.

UR - https://www.scopus.com/pages/publications/85169763342

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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