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Related Experiment Video

Updated: Jan 23, 2026

In Situ Characterization of Boehmite Particles in Water Using Liquid SEM
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SEM/FIB Imaging for Studying Neural Interfaces.

Itai Henn1,2, Ayelet Atkins1,2, Amos Markus1

  • 1Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat Gan, Israel.

Developmental Neurobiology
|June 23, 2019
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Summary
This summary is machine-generated.

Optimizing sample preparation for scanning electron microscopy is crucial for studying cell interfaces. Resin drying and OTOTO staining best preserve cell morphology and structure for regenerative therapies.

Keywords:
3D scaffoldsSEM/FIBcellular membraneelectron microscopefixationinterfacestaining

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Area of Science:

  • Tissue and neural engineering
  • Regenerative medicine
  • Biomedical applications

Background:

  • Studying cell-material interfaces is vital for regenerative therapies.
  • Scanning electron microscopy with focused ion beam offers high resolution but requires optimized sample preparation.
  • Preserving the natural cell structure during fixation, dehydration, and staining is challenging.

Purpose of the Study:

  • To compare and optimize sample drying and staining protocols for scanning electron microscopy.
  • To preserve cells in a life-like state for studying cell interfaces with 3D structures and electrodes.
  • To determine the best methods for visualizing intracellular organelles and membranes.

Main Methods:

  • Chemical fixation using glutaraldehyde and formaldehyde.
  • Comparison of four drying techniques: critical point drying, hexamethyldisiloxane, osmium tetroxide-thiocarbohydrazide (OTOTO), and resin.
  • Evaluation of four staining techniques: osmium tetroxide, osmium tetroxide and salts, osmium and uranyl acetate, and OTOTO.
  • Experiments conducted on photoreceptor precursors, neural cells, and retinal pigment epithelial cells.

Main Results:

  • Resin drying combined with OTOTO staining emerged as the optimal processing combination.
  • This combination demonstrated excellent preservation of cell morphology and cell interface.
  • It resulted in the lowest percentage of cellular protrusion breakage and high-quality imaging.

Conclusions:

  • The optimized protocol enhances the understanding of cell-structure interfaces.
  • This advancement is expected to improve various biomedical applications in regenerative medicine.
  • The findings provide a foundation for future research in tissue and neural engineering.