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Updated: Aug 24, 2025
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3D Imaging of Soft-Tissue Samples using an X-ray Specific Staining Method and Nanoscopic Computed Tomography
Published on: October 24, 2019
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In situ X-ray-assisted electron microscopy staining for large biological samples
Sebastian Ströh1, Eric W Hammerschmith1, David W Tank1
1Princeton Neuroscience Institute, Princeton University, Princeton, United States.
Elife
|October 20, 2022
Summary
This study introduces X-ray-assisted staining for real-time observation of electron microscopy staining processes. This method enables efficient optimization of staining protocols for large biological samples like entire brains.
Area of Science:
- Biological imaging
- Materials science
Background:
- Electron microscopy enables ultrastructural analysis of large biological samples.
- Homogeneous staining of large samples remains a significant challenge.
- Current staining quality assessment is time-consuming and inefficient for large samples.
Purpose of the Study:
- To develop a method for real-time observation of electron microscopy staining.
- To optimize staining protocols for large biological samples.
- To understand heavy metal accumulation dynamics during staining.
Main Methods:
- Developed an in situ time-lapsed X-ray-assisted staining procedure.
- Measured heavy metal (osmium) accumulation in large tissue samples.
- Investigated tissue volume changes in different staining solutions.
- Developed a diffusion-reaction-advection model for osmium accumulation.
Main Results:
- Osmium accumulation in fixed tissue follows a quadratic dependence on incubation time and sample size.
- Potassium ferrocyanide clears tissue after osmium staining.
- Tissue expands in osmium tetroxide and shrinks in reduced osmium solutions.
- The developed model accurately simulates osmium accumulation.
Conclusions:
- X-ray-assisted staining provides real-time insights into staining kinetics.
- This method facilitates simulation-guided optimization of staining protocols.
- Enables reliable staining procedures for large samples like whole brains.
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