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Preparation of Samples for Electron Microscopy

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To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...

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Updated: Dec 25, 2025

Multimodal Hierarchical Imaging of Serial Sections for Finding Specific Cellular Targets within Large Volumes

Multimodal Hierarchical Imaging of Serial Sections for Finding Specific Cellular Targets within Large Volumes

Published on: March 20, 2018

Surface cleaning and sample carrier for complementary high-resolution imaging techniques.

Pietro Benettoni¹, Jia-Yu Ye¹, Timothy R Holbrook²

¹Department of Isotope Biogeochemistry, Helmholtz Centre for Environmental Research-UFZ, Permoserstraße 15, 04318 Leipzig, Germany.

|March 28, 2020

Summary

This summary is machine-generated.

A novel hydrophilic polymer-coated silicon wafer sample carrier was developed for advanced imaging. This carrier minimizes the coffee-ring effect, preserving analyte integrity for diverse high-resolution techniques.

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Area of Science:

Materials Science
Analytical Chemistry
Microscopy

Background:

High-resolution imaging techniques require complementary analytical approaches.
A common sample carrier is needed for diverse preparation methods and analytes.
Existing carriers struggle to maintain analyte composition and arrangement during preparation.

Purpose of the Study:

To develop a new sample carrier for complementary high-resolution imaging.
To ensure analyte integrity, pristine composition, and arrangement during sample preparation.
To overcome limitations of existing sample carriers in multi-modal analysis.

Main Methods:

Development of a silicon wafer with a robust, hydrophilic polymer coating.
Cross-linking and stoving to enhance solvent resistance of the polymer.
UV-ozone cleaning for improved adhesion, reproducible surface properties, and hydrophilicity recovery.
Characterization using 3D optical profilometry and contact angle measurements.
Testing with nanoparticles, bacterial cells, and tissue sections.
Analysis using optical microscopy, scanning electron microscopy, helium ion microscopy, laser ablation inductively coupled plasma mass spectrometry, and time-of-flight secondary ion mass spectrometry.

Main Results:

A novel, robust hydrophilic polymer-coated silicon wafer sample carrier was successfully developed.
The carrier exhibits spongelike behavior, absorbing solvent and solutes to prevent the coffee-ring effect.
Analyte deposition shows enhanced homogeneity and preservation of native identity.
The carrier demonstrated suitability for diverse sample types including nanoparticles, bacteria, and tissue.

Conclusions:

The developed sample carrier is highly effective for complementary high-resolution imaging.
Its unique properties enable precise analyte deposition and preservation of integrity.
This innovation offers broad applicability and significant value for multi-modal analytical workflows.