string(19) "en/achievements/742" Step-by-step procedure for an optimized serial lift-out cryo-focused ion beam milling technique in tissue analysis-Shenzhen Medical Academy of Research and Translation
Home Achievements

Step-by-step procedure for an optimized serial lift-out cryo-focused ion beam milling technique in tissue analysis

Date：2022/02/09

Source：深圳医学科学院

This paper presents an optimized serial lift-out cryo-focused ion beam milling technique for tissue samples. By incorporating a six-sided attachment geometry and adjustable grid trench configurations, this method significantly enhances the mechanical stability and reproducibility of lamellae while reducing operator dependency. Using mouse intestinal tissue as a model, combined with high-pressure freezing and cryo-CLEM localization, the study successfully prepared high-quality lamellae suitable for cryo-electron tomography, clearly resolving microvillar membranes, actin bundles, and nanobristle-like structures on the membrane surface. The Structural Biology Core Facility provided critical equipment and technical support for this study: the Helios 5 Hydra CX plasma cryo-dual beam microscope was used for milling and lamella preparation; platform engineers were responsible for tomographic tilt series data collection and TEM maintenance; data processing relied on the Amira software deployed at the facility for 3D segmentation and rendering. The facility's cryo-EM, CLEM correlation system, and professional technical personnel provided full-process support for in situ tissue structure analysis.

 

Back List
Prev：Structural plasticity of mouse intestinal microvilli revealed by tissue-level cryo-ET Next：Evolutionary repurposing of a metabolic thiolase complex enables antibiotic biosynthesis
Message