TY  - INPR
A1  - Tuijtel, Maarten W.
A1  - Kreysing, Jan Philipp
A1  - Welsch, Sonja
A1  - Hummer, Gerhard
A1  - Beck, Martin
A1  - Turoňová, Beata
T1  - Thinner is not always better: Optimising cryo lamellae for subtomogram averaging
T2  - bioRxiv
N2  - Cryo-electron tomography (cryo-ET) is a powerful method to elucidate subcellular architecture and to structurally analyse biomolecules in situ by subtomogram averaging (STA). Specimen thickness is a key factor affecting cryo-ET data quality. Cells that are too thick for transmission imaging can be thinned by cryo-focused-ion-beam (cryo-FIB) milling. However, optimal specimen thickness for cryo-ET on lamellae has not been systematically investigated. Furthermore, the ions used to ablate material can cause damage in the lamellae, thereby reducing STA resolution. Here, we systematically benchmark the resolution depending on lamella thickness and the depth of the particles within the sample. Up to ca. 180 nm, lamella thickness does not negatively impact resolution. This shows that there is no need to generate very thin lamellae and thickness can be chosen such that it captures major cellular features. Furthermore, we show that gallium-ion-induced damage extends to depths of up to 30 nm from either lamella surface.
Y1  - 2023
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/75088
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-750881
IS  - 2023.07.31.551274
ER  -