English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  A cryo-FIB lift-out technique enables molecular-resolution cryo-ET within native Caenorhabditis elegans tissue

Schaffer, M., Pfeffer, S., Mahamid, J., Kleindiek, S., Laugks, T., Albert, S., et al. (2019). A cryo-FIB lift-out technique enables molecular-resolution cryo-ET within native Caenorhabditis elegans tissue. Nature Methods, 16(8), 757-762. doi:10.1038/s41592-019-0497-5.

Item is

Basic

show hide
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Schaffer, Miroslava1, Author           
Pfeffer, Stefan2, Author
Mahamid, Julia2, Author
Kleindiek, Stephan2, Author
Laugks, Tim1, Author           
Albert, Sahradha1, Author           
Engel, Benjamin D.1, Author           
Rummel, Andreas2, Author
Smith, Andrew J.2, Author
Baumeister, Wolfgang1, Author           
Plitzko, Juergen M.1, Author           
Affiliations:
1Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              
2external, ou_persistent22              

Content

show
hide
Free keywords: SAMPLE PREPARATION; CRYOELECTRON TOMOGRAPHY; BEAM; CELLS; SOCIOLOGY; STEMBiochemistry & Molecular Biology;
 Abstract: Cryo-focused ion beam milling of frozen-hydrated cells has recently provided unprecedented insights into the inner space of cells. In combination with cryo-electron tomography, this method allows access to native structures deep inside cells, enabling structural studies of macromolecules in situ. However, this approach has been mainly limited to individual cells that can be completely vitrified by plunge-freezing. Here, we describe a preparation method that is based on the targeted extraction of material from high-pressure-frozen bulk specimens with a cryo-gripper tool. This lift-out technique enables cryo-electron tomography to be performed on multicellular organisms and tissue, extending the range of applications for in situ structural biology. We demonstrate the potential of the lift-out technique with a structural study of cytosolic 80S ribosomes in a Caenorhabditis elegans worm. The preparation quality allowed for subtomogram analysis with sufficient resolution to distinguish individual ribosomal translocation states and revealed significant cell-to-cell variation in ribosome structure.

Details

show
hide
Language(s): eng - English
 Dates: 2019
 Publication Status: Published in print
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000477857700033
DOI: 10.1038/s41592-019-0497-5
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Nature Methods
  Other : Nature Methods
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: New York, NY : Nature Pub. Group
Pages: - Volume / Issue: 16 (8) Sequence Number: - Start / End Page: 757 - 762 Identifier: ISSN: 1548-7091
CoNE: https://pure.mpg.de/cone/journals/resource/111088195279556