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

No Thumbnail Available
File version
Author(s)
Ashtiani, Dariush
Venugopal, Hari
Belousoff, Matthew
Spicer, Bradley
Mak, Johnson
Neild, Adrian
de Marco, Alex
Griffith University Author(s)
Primary Supervisor
Other Supervisors
Editor(s)
Date
2018
Size
File type(s)
Location
License
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.

Journal Title

Journal of Structural Biology

Conference Title
Book Title
Edition
Volume

203

Issue

2

Thesis Type
Degree Program
School
Publisher link
Patent number
Funder(s)
Grant identifier(s)
Rights Statement
Rights Statement
Item Access Status
Note
Access the data
Related item(s)
Subject

Biochemistry and cell biology

Biochemistry and cell biology not elsewhere classified

Zoology

Persistent link to this record
Citation
Collections