Targeted imaging of specialized plant cell walls by improved cryo-CLEM and cryo-electron tomography.

ABSTRACT: Cryo-focused ion beam scanning electron microscopy (Cryo-FIBSEM) has become essential for preparing cryo-lamellae. Here, we present a series of improvements that speed up and enhance the efficiency of the Serial Lift-Out and SOLIST (Serialized On-grid Lift-In Sectioning for Tomography) procedures. We extend the cryo-FIBSEM session to 5 days and eliminate the need of copper or gold block intermediates and reduce curtaining effects. Finally, we report a routine to target lamellae with a precision of approximately 1 µm in X/Y/Z. We demonstrate the power of our improvements by targeting Casparian strips, suberin lamellae, as well as xylem vessels of Arabidopsis thaliana roots. This requires reaching a target of 5 micrometers in a 3 mm long and 80-120 µm thick root section. Despite ice formatio

SUBMITTER: Daraspe J 

PROVIDER: S-EPMC12728224 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature