If you find this useful, please cite our work.
There is a bioRxiv preprint available at: https://doi.org/10.1101/797506
Genevieve Buckley, Gediminas Gervinskas, Cyntia Taveneau, Hari Venugopal, James C. Whisstock, Alex de Marco. Automated cryo-lamella preparation for high-throughput in-situ structural biology. bioRxiv, 2019.
BibTex:
@article {Buckley797506,
author = {Buckley, Genevieve and Gervinskas, Gediminas and Taveneau, Cyntia and Venugopal, Hari and Whisstock, James C. and de Marco, Alex},
title = {Automated cryo-lamella preparation for high-throughput in-situ structural biology},
elocation-id = {797506},
year = {2019},
doi = {10.1101/797506},
publisher = {Cold Spring Harbor Laboratory},
abstract = {Cryo-transmission electron tomography (cryo-ET) in association with cryo-focused ion beam (cryo-FIB) milling enables structural biology studies to be performed directly within the cellular environment. Cryo-preserved cells are milled and a lamella with a thickness of 200-300 nm provides an electron transparent window suitable for cryo-ET imaging. Cryo-FIB milling is an effective method, but it is a tedious and time-consuming process, which typically results in ~10 lamellae per day. Here, we introduce an automated method to reproducibly prepare cryo-lamellae on a grid and reduce the amount of human supervision. We tested the routine on cryo-preserved Saccharomyces cerevisiae and demonstrate that this method allows an increased throughput, achieving a rate of 5 lamellae/hour without the need to supervise the FIB milling. We demonstrate that the quality of the lamellae is consistent throughout the preparation and their compatibility with cryo-ET analyses.},
URL = {https://www.biorxiv.org/content/early/2019/10/08/797506},
eprint = {https://www.biorxiv.org/content/early/2019/10/08/797506.full.pdf},
journal = {bioRxiv}
}