Related Experiment Video
Updated: Jul 9, 2026
Chronic Imaging of Mouse Visual Cortex Using a Thinned-skull Preparation
Published on: October 25, 2010
Cryo-Electron Tomography Workflow for Thick Tissues Demonstrated in Mouse Hippocampus
Jinrong Hu1, Xiaokang Zhang2, Jing Wu3
1Cryo-EM Center, Southern University of Science & Technology.
A new workflow enables 3D visualization of cellular structures in thick tissues using cryo-electron tomography (cryo-ET). This method improves sample preparation and data acquisition for high-throughput structural biology studies.
Area of Science:
- Structural Biology
- Cell Biology
- Microscopy
Background:
- Cryo-electron tomography (cryo-ET) offers 3D visualization of cellular ultrastructure.
- Thick tissue application is hindered by vitrification and lamella preparation challenges.
Purpose of the Study:
- To present an end-to-end workflow for cryo-ET of thick tissues.
- To enable high-throughput, in situ structural biology studies.
Main Methods:
- Integrated workflow: high-pressure freezing (HPF), cryo-FIB lift-out, controlled lamella thinning (120-200 nm).
- Standardized conductive coating to minimize curtaining.
- Automated, dose-symmetric data acquisition (PACEtomo scripts) and near-real-time processing (AreTomo 3).
Main Results:
- Routine production of well-vitrified samples and uniform lamellae (150 ± 40 nm).
- High-throughput tilt-series collection (5-8 min/series).
- Demonstrated on mouse hippocampus.
Conclusions:
- The workflow provides a reproducible method for cryo-ET of complex tissues.
- Enables direct study of in situ structural biology.
- Overcomes limitations of previous cryo-ET methods for thick samples.
More Related Videos
09:59Preparation of Primary Neurons for Visualizing Neurites in a Frozen-hydrated State Using Cryo-Electron Tomography
Published on: February 12, 2014
11:20Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03YEMK
Published on: December 19, 2019
