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Imaging Biological Samples with Optical Microscopy

Imaging Biological Samples with Optical Microscopy

Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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Related Experiment Video

Updated: Jul 2, 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

Beam spreading and spatial resolution in thick organic specimens.

Jerome K Hyun¹, Peter Ercius, David A Muller

¹Department of Physics, Cornell University, E13 Clark Hall, Ithaca, NY 14853, USA. jkh32@cornell.edu

Ultramicroscopy

|August 30, 2008

Summary

Scanning transmission electron microscopy (STEM) tomography enables 3D imaging of thick biological samples. Beam divergence, not scattering, limits resolution in polymers over 100 nm, so minimizing divergence enhances spatial resolution.

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Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography

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Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography

Published on: March 12, 2017

Area of Science:

Electron Microscopy
Materials Science
Biophysics

Background:

Scanning transmission electron microscopy (STEM) tomography is a powerful technique for 3D imaging.
Image resolution in STEM tomography is often limited by electron scattering within the sample.
Understanding the interplay between material thickness and spatial resolution is crucial for optimizing imaging.

Purpose of the Study:

To investigate the factors limiting spatial resolution in STEM tomography of micron-thick biological specimens and nanostructures.
To analyze the competition between beam divergence and beam spreading due to plural elastic scattering.
To determine the optimal conditions for enhanced spatial resolution in STEM imaging.

Main Methods:

Utilized scanning transmission electron microscopy (STEM) for tomographic imaging.
Analyzed the effects of material thickness on spatial resolution.
Quantified the contributions of beam divergence and beam spreading (plural elastic scattering).

Main Results:

Beam divergence was found to dominate beam spreading in typical embedding polymers thicker than 100 nm.
Minimizing beam divergence was shown to significantly enhance spatial resolution.
The challenges associated with resolution limitations are amplified in spherical-aberration-corrected instruments due to their shorter depth of field.

Conclusions:

Spatial resolution in STEM tomography of thicker samples is primarily governed by beam divergence rather than plural elastic scattering.
Optimizing probe characteristics to minimize beam divergence is key to achieving higher resolution.
Further research is needed to fully exploit STEM tomography for thick biological and nanostructure imaging.