English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  A streamlined workflow for automated cryo focused ion beam milling

Tacke, S., Erdmann, P., Wang, Z., Klumpe, S., Grange, M., Plitzko, J., et al. (2021). A streamlined workflow for automated cryo focused ion beam milling. Journal of Structural Biology, 213(3): 107743. doi:10.1016/j.jsb.2021.107743.

Item is

Basic

show hide
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Tacke, Sebastian1, Author
Erdmann, Philipp2, Author              
Wang, Zhexin1, Author
Klumpe, Sven2, Author              
Grange, Michael1, Author
Plitzko, Jürgen2, Author              
Raunser, Stefan1, Author
Affiliations:
1external, ou_persistent22              
2Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

Content

show
hide
Free keywords: VITREOUS SECTIONS; CRYOELECTRON; TOMOGRAPHYBiochemistry & Molecular Biology; Biophysics; Cell Biology; Electron tomography; Focused ion beam; Cryo shield; Cryo shutter; Contamination; Automation;
 Abstract: Cryo-electron tomography (cryo-ET) is an emerging technique to study the cellular architecture and the structure of proteins at high resolution in situ. Most biological specimens are too thick to be directly investigated and are therefore thinned by milling with a focused ion beam under cryogenic conditions (cryo-FIB). This procedure is prone to contaminations, which makes it a tedious process, often leading to suboptimal results. Here, we present new hardware that overcomes the current limitations. We developed a new glove box and a high vacuum cryo transfer system and installed a stage heater, a cryo-shield and a cryo-shutter in the FIB milling microscope. This reduces the ice contamination during the transfer and milling process and simplifies the handling of the sample. In addition, we tested a new software application that automates the key milling steps. Together, these improvements allow for high-quality, high-throughput cryo-FIB milling. This paves the way for new types of experiments, which have been previously considered infeasible.

Details

show
hide
Language(s): eng - English
 Dates: 2021
 Publication Status: Published online
 Pages: 10
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000692079400005
DOI: 10.1016/j.jsb.2021.107743
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Journal of Structural Biology
  Abbreviation : J. Struct. Biol.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: San Diego, CA : Elsevier
Pages: - Volume / Issue: 213 (3) Sequence Number: 107743 Start / End Page: - Identifier: ISSN: 1047-8477
CoNE: https://pure.mpg.de/cone/journals/resource/954922650160