If you find this useful, please cite our work.
Genevieve Buckley, Gediminas Gervinskas, Cyntia Taveneau, Hariprasad Venugopal, James C. Whisstock, Alex de Marco, Automated cryo-lamella preparation for high-throughput in-situ structural biology, Journal of Structural Biology, Volume 210, Issue 2, 2020 https://doi.org/10.1016/j.jsb.2020.107488.
BibTex:
@article{BUCKLEY2020107488,
title = "Automated cryo-lamella preparation for high-throughput in-situ structural biology",
journal = "Journal of Structural Biology",
volume = "210",
number = "2",
pages = "107488",
year = "2020",
issn = "1047-8477",
doi = "https://doi.org/10.1016/j.jsb.2020.107488",
url = "http://www.sciencedirect.com/science/article/pii/S104784772030054X",
author = "Genevieve Buckley and Gediminas Gervinskas and Cyntia Taveneau and Hariprasad Venugopal and James C. Whisstock and Alex {de Marco}",
keywords = "Cryo-FIB, Cryo-lamella, Automation, cryo-EM, In situ structural biology",
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 typical 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, mammalian 293 T cells, and lysozyme protein crystals. Here we demonstrate that our 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."
}
You can download a PDF of our published work at: https://doi.org/10.1016/j.jsb.2020.107488
There is also a bioRxiv preprint of this work available at: https://doi.org/10.1101/797506