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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

Published on: March 12, 2017

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Micro-CT scouting for transmission electron microscopy of human tissue specimens

A G Morales1, E S Stempinski1, X Xiao2

  • 1National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, U.S.A.

Journal of Microscopy
|February 9, 2016
PubMed

Insights

Micro-computed tomography (micro-CT) scouting noninvasively identifies regions of interest in tissue samples for transmission electron microscopy (TEM). This method significantly reduces sectioning time and labor for pathology studies.

Area of Science:

  • Materials Science
  • Biomedical Engineering
  • Pathology

Background:

  • Transmission electron microscopy (TEM) offers sub-nanometre resolution for volumetric samples.
  • Metal staining enhances contrast in pathology specimens but renders them opaque to light microscopy.
  • Locating specific regions of interest within stained samples for TEM is a time-consuming process involving extensive sectioning.

Purpose of the Study:

  • To introduce and evaluate micro-computed tomography (micro-CT) scouting as a method for noninvasively identifying regions of interest in samples for TEM.
  • To streamline the process of preparing tissue samples for high-resolution ultrastructural analysis using TEM.

Main Methods:

  • Utilized a bench-top micro-CT scanner with 10 micrometre resolution for sample scouting.
  • Applied micro-CT to identify mucous membrane patches in osmium-stained human nasal scraping samples.
  • Guided ultra-thin sectioning for TEM based on micro-CT identified regions of interest.

Main Results:

  • Successfully located regions of interest within osmium-stained human nasal tissue using micro-CT.
  • Reduced the number of sections required for analysis from approximately 20 (light microscopy) to three.
  • Enabled direct TEM inspection of cilia in membrane epithelial cells with nanometre resolution.

Conclusions:

  • Micro-CT scouting is an effective noninvasive technique for guiding sample sectioning for TEM.
  • This approach significantly decreases the time and labor associated with preparing pathology samples for high-resolution microscopy.
  • The method facilitates efficient identification and analysis of ultrastructural details in biological tissues.
Keywords:
bench-top micro-CT scannermicro-CT scoutingthree-dimensional visualizationtransmission electron microscopy

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