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Related Concept Videos

Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...

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Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass
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Quantifying Golgi structure using EM: combining volume-SEM and stereology for higher throughput.

Sophie Ferguson1, Anna M Steyer2, Terry M Mayhew3

  • 1Structural Cell Biology Group, School of Medicine, University of St Andrews, North Haugh, Fife, KY16 9TF, Scotland, UK.

Histochemistry and Cell Biology
|April 22, 2017
PubMed
Summary

Sampling-based stereology significantly reduces data and workload for high-throughput Golgi complex analysis using volume scanning electron microscopy (volume-SEM). This method enables accurate quantitative structural data from fewer images, enhancing efficiency in cell biology research.

Keywords:
FIBSEMGolgiQuantificationSBF-SEMSamplingStereologyVolume-SEM

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Last Updated: Jun 27, 2026

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Quantitative Morphology

Background:

  • High-throughput quantitative morphological analysis of organelles like the Golgi complex is crucial for understanding cellular responses.
  • Light microscopy (LM) lacks resolution for fine Golgi structures, while traditional transmission electron microscopy (TEM) is time-consuming.
  • Volume scanning electron microscopy (volume-SEM) offers high resolution but generates large datasets requiring extensive 3D reconstruction.

Purpose of the Study:

  • To reduce the data storage, digital waste, and workload associated with volume-SEM analyses.
  • To demonstrate a method for efficient quantitative analysis of Golgi complex morphology using volume-SEM.
  • To adapt sampling-based stereology for high-throughput organelle analysis.

Main Methods:

  • Application of sampling-based stereology to volume-SEM data.
  • Quantitative sensing of Golgi populations using single random slices.
  • Acquisition of structural data from individual cells using 5-10 volume-SEM sections.

Main Results:

  • Substantial reduction in information storage, digital waste, and workload for volume-SEM.
  • Accurate quantitative structural data on Golgi organelles obtained from a limited number of sections.
  • Successful sensing of population parameters and cell-cell variability in Golgi structures.

Conclusions:

  • Sampling-based stereology offers a more efficient approach for quantitative Golgi analysis with volume-SEM.
  • This method enhances the speed and relevance of data acquisition in volume-SEM.
  • The approach is particularly useful for techniques like correlative LM and EM (CLEM) with variable cell responses.