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Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy
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Work smart, not hard: How array tomography can help increase the ultrastructure data output.

Irina Kolotuev1

  • 1Electron Microscopy Facility, University of Lausanne, Lausanne, Switzerland.

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|August 26, 2023
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Summary

Array Tomography (AT) offers versatile ultrastructure analysis for biological samples, overcoming Transmission Electron Microscopy (TEM) limitations. This method enables detailed imaging for research and diagnostics.

Keywords:
SEMarray tomographyblood cellsdiagnostic vEMhistologymodel organismsvolume electron microscopy

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biotechnology

Background:

  • Transmission electron microscopy (TEM) has been a cornerstone of cell biology for decades.
  • Advancements in volume electron microscopy, like serial block face and focused ion beam scanning electron microscopy, have enhanced ultrastructure analysis.
  • Array Tomography (AT) emerged as a sequential imaging technique for resin-embedded samples.

Purpose of the Study:

  • To highlight the capabilities and versatility of Array Tomography (AT).
  • To demonstrate AT's potential in overcoming limitations of traditional Transmission Electron Microscopy (TEM).
  • To showcase AT's applicability in various biological research and diagnostic contexts.

Main Methods:

  • Sequential image acquisition of resin-embedded sample sections on a large support.
  • Immunolabelling with multiple fluorescent labels, combined with ultrastructure observation.
  • Generating and imaging serial sections for 3D reconstruction using scanning electron microscopy (SEM).

Main Results:

  • AT facilitates tasks difficult or impossible with conventional TEM.
  • The technique is applicable to diverse biological samples for selected sections or volume analysis.
  • AT is compatible with morphological analysis, multiplex immunolabelling, immune-gold labelling, and correlative light and electron microscopy.

Conclusions:

  • Array Tomography (AT) presents a versatile and straightforward method for ultrastructure analysis.
  • Its compatibility with multiple labelling techniques and workflows makes it valuable for basic research.
  • The technique's adaptability positions it as a potential diagnostic tool with a simplified routine.