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Related Experiment Video

Updated: May 13, 2026

Serial Block-Face Scanning Electron Microscopy (SBF-SEM) of Biological Tissue Samples

Serial Block-Face Scanning Electron Microscopy (SBF-SEM) of Biological Tissue Samples

Published on: March 26, 2021

Automated in-chamber specimen coating for serial block-face electron microscopy.

B Titze¹, W Denk¹

¹Department of Biomedical Optics, Max Planck Institute for Medical Research, Heidelberg, Germany.

Journal of Microscopy

|March 5, 2013

Summary

This summary is machine-generated.

This study introduces an in-situ coating method for scanning electron microscopy (SEM) to prevent sample charging in serial block-face electron microscopy (SBEM). This technique enhances image quality and enables new imaging modes for insulating specimens.

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Area of Science:

Materials Science
Microscopy Techniques
Nanotechnology

Background:

Insulating specimens in scanning electron microscopy (SEM) suffer from sample charging, distorting images.
Traditional conductive coating methods are incompatible with serial block-face electron microscopy (SBEM) due to the in-situ cutting and imaging process.

Purpose of the Study:

To develop a method to eliminate charging effects in SBEM for insulating samples.
To improve image quality and enable advanced imaging modes in SBEM.

Main Methods:

Integrated an electron beam evaporator into the SEM chamber for in-situ metallic film coating (1-2 nm) after each cutting cycle.
Automated the coating process for serial acquisition of SBEM stacks.
Utilized beam deceleration for enhanced contrast on non-conducting samples.

Main Results:

Successfully eliminated charging effects for both backscattered (BSE) and secondary electron (SE) imaging on samples up to 12 mm.
Observed minimal signal-to-noise ratio (SNR) reduction compared to low-vacuum methods.
Demonstrated acquisition of over 1000 cut/coat/image cycles and successful imaging using beam deceleration and SE contrast.

Conclusions:

In-situ coating is a viable solution for charging artifacts in SBEM.
This method preserves sample integrity and enhances imaging capabilities for insulating materials.
The automated process facilitates high-resolution 3D reconstructions of challenging specimens.