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Preparation of Samples for Electron Microscopy

Preparation of Samples for Electron Microscopy

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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Related Experiment Video

Updated: Sep 17, 2025

Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy

Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy

Published on: February 5, 2017

Enabling Liquid Scanning Electron Microscopy for Therapeutic Suspensions Using Vacuum-Compatible Liquid Capsules.

Raina Borum¹, Paul Reichert², Zhao Li³

¹Analytical Research and Development, Merck & Co., Inc., Rahway, New Jersey 07065, United States.

Analytical Chemistry

|July 3, 2025

Summary

This summary is machine-generated.

We enhanced liquid scanning electron microscopy (wet SEM) for imaging therapeutic suspensions by improving sample adhesion and contrast. This allows for high-resolution, in situ analysis of small organic and biologic materials, aiding drug development.

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Last Updated: Sep 17, 2025

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Area of Science:

Materials Science
Biotechnology
Analytical Chemistry

Background:

Liquid scanning electron microscopy (wet SEM) enables high-resolution imaging of materials in liquid environments.
Challenges like poor sample adhesion, radiation sensitivity, and low contrast limit wet SEM for therapeutic suspensions.
Existing methods are insufficient for characterizing small organic and biologic materials in native states.

Purpose of the Study:

To overcome limitations of wet SEM for analyzing therapeutic suspensions.
To expand the application of wet SEM for in situ characterization of small organic and biologic materials.
To improve resolution and contrast for imaging submicrometer particles in suspension.

Main Methods:

Utilized a vacuum-compatible liquid cell with surface-modified imaging windows for enhanced sample adhesion.
Employed imaging spacers to improve sampling efficiency for heterogeneous particle sizes.
Incorporated gadolinium acetate to enhance contrast and resolution via Z-contrast.

Main Results:

Demonstrated significantly improved sample adhesion on surface-coated imaging windows compared to noncoated ones.
Achieved a resolution of approximately 10 nm for submicrometer protein particles in suspension.
Successfully elucidated particle attribute details influencing rheological and pharmacokinetic behaviors in situ.

Conclusions:

Developed strategies to overcome key limitations of wet SEM for therapeutic suspensions.
Established wet SEM as a versatile tool for in situ analysis of organic and biologic soft materials.
Highlighted the importance of high-resolution imaging for advancing therapeutic process development and formulation design.