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Updated: Feb 24, 2026

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Correlative scanning electron and super-resolution structured illumination microscopy.

Joseph R Hamiliton1, Summer K Levis1, Guy M Hagen1

  • 1UCCS BioFrontiers Center, University of Colorado Colorado Springs, 1420 Austin Bluffs Parkway, Colorado Springs, Colorado, 80918, USA.

Biorxiv : the Preprint Server for Biology
|February 23, 2026
PubMed
Summary
This summary is machine-generated.

Correlative light and electron microscopy (CLEM) enhances biological sample analysis. Scanning electron microscopy (SEM) provides higher resolution and validates results from super-resolution structured illumination fluorescence microscopy (SIM).

Keywords:
CLEMNanoSuitSEMSIMSSEScanning electron microscopyStructured illumination microscopycorrelative light and electron microscopysuper-resolutionsurface shield enhancer

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Area of Science:

  • Biological sciences
  • Microscopy
  • Cell biology

Background:

  • Correlative microscopy integrates multiple imaging techniques for comprehensive sample analysis.
  • Preservation methods are crucial for preparing biological samples for correlative light and electron microscopy (CLEM).

Purpose of the Study:

  • To compare the image quality and resolution of widefield (WF) microscopy, super-resolution structured illumination fluorescence microscopy (SIM), and scanning electron microscopy (SEM) in a correlative workflow.
  • To evaluate the effectiveness of NanoSuit chemical treatment in preserving sample integrity for SEM after fluorescence imaging.

Main Methods:

  • A mammalian testis sample was imaged using WF and SIM fluorescence microscopy.
  • The sample underwent NanoSuit chemical treatment.
  • Post-treatment, the sample was imaged using SEM.

Main Results:

  • SEM achieved higher resolution compared to WF and SIM.
  • SEM provided structural details that validated the findings from SIM.
  • NanoSuit treatment enabled successful SEM imaging after fluorescence microscopy.

Conclusions:

  • CLEM, particularly combining SIM and SEM, offers enhanced insights into biological sample ultrastructure.
  • SEM is a valuable tool for validating high-resolution fluorescence microscopy data.
  • The NanoSuit protocol is effective for preparing samples for correlative SEM analysis.