Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation

Dariush Ashtiani, Hari Venugopal, Matthew Belousoff, Bradley Spicer, Johnson Mak, Adrian Neild, Alex de Marco

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6–8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves.

Original languageEnglish
Pages (from-to)94-101
Number of pages8
JournalJournal of Structural Biology
Volume203
Issue number2
DOIs
Publication statusPublished - Aug 2018

Keywords

  • Atomization
  • Blot-free sample preparation
  • Cryo-EM
  • Microfluidics
  • Surface acoustic waves

Cite this

@article{1c176b41df674371828ef072f0e01628,
title = "Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation",
abstract = "Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6–8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves.",
keywords = "Atomization, Blot-free sample preparation, Cryo-EM, Microfluidics, Surface acoustic waves",
author = "Dariush Ashtiani and Hari Venugopal and Matthew Belousoff and Bradley Spicer and Johnson Mak and Adrian Neild and {de Marco}, Alex",
year = "2018",
month = "8",
doi = "10.1016/j.jsb.2018.03.012",
language = "English",
volume = "203",
pages = "94--101",
journal = "Journal of Structural Biology",
issn = "1047-8477",
publisher = "Elsevier",
number = "2",

}

Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation. / Ashtiani, Dariush; Venugopal, Hari; Belousoff, Matthew; Spicer, Bradley; Mak, Johnson; Neild, Adrian; de Marco, Alex.

In: Journal of Structural Biology, Vol. 203, No. 2, 08.2018, p. 94-101.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation

AU - Ashtiani, Dariush

AU - Venugopal, Hari

AU - Belousoff, Matthew

AU - Spicer, Bradley

AU - Mak, Johnson

AU - Neild, Adrian

AU - de Marco, Alex

PY - 2018/8

Y1 - 2018/8

N2 - Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6–8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves.

AB - Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6–8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves.

KW - Atomization

KW - Blot-free sample preparation

KW - Cryo-EM

KW - Microfluidics

KW - Surface acoustic waves

UR - http://www.scopus.com/inward/record.url?scp=85046163816&partnerID=8YFLogxK

U2 - 10.1016/j.jsb.2018.03.012

DO - 10.1016/j.jsb.2018.03.012

M3 - Article

VL - 203

SP - 94

EP - 101

JO - Journal of Structural Biology

JF - Journal of Structural Biology

SN - 1047-8477

IS - 2

ER -