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Related Experiment Video

Updated: May 15, 2025

Use of Micro X-ray Computed Tomography with Phosphotungstic Acid Preparation to Visualize Human Fibromuscular Tissue
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Phosphotungstic Acid Staining to Visualize the Vagus Nerve Perineurium Using Micro-CT.

Aniruddha R Upadhye1,2, Eleana Cintron1, Jichu Zhang1

  • 1Department of Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio, USA.

Journal of Neuroimaging : Official Journal of the American Society of Neuroimaging
|April 10, 2025
PubMed
Summary

Phosphotungstic acid (PTA) effectively visualizes peripheral nerve structures using micro-CT imaging. This contrast agent is compatible with downstream histology, aiding the development of selective nerve therapies.

Keywords:
X‐ray microtomographyneuroanatomyperipheral nervesphosphotungstic acidvagus nerve

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Area of Science:

  • Neuroscience
  • Medical Imaging
  • Histology

Background:

  • Peripheral nerve stimulation offers therapeutic potential but is limited by side effects.
  • Understanding nerve fascicular and fiber organization is crucial for developing selective therapies.
  • Micro-CT imaging of contrast-stained nerves can visualize microstructures like perineurium and vasculature.

Purpose of the Study:

  • To evaluate phosphotungstic acid (PTA) as a micro-CT contrast agent for nerve imaging.
  • To assess the compatibility of PTA staining with downstream histological analysis.
  • To optimize micro-CT parameters for enhanced nerve visualization.

Main Methods:

  • Human vagus nerve samples were stained with Lugol's iodine, osmium tetroxide, and PTA.
  • Micro-CT imaging was performed, and contrast ratios of nerve microstructures were quantified.
  • PTA-stained samples were processed for histology (H&E, Masson's trichrome, IHC) to assess tissue integrity.

Main Results:

  • PTA provided superior visualization of the perineurium compared to iodine and osmium tetroxide.
  • Optimized micro-CT parameters (55 kV, 109 µA) balanced noise and sharpness for PTA-stained nerves.
  • PTA was generally nondestructive for histology, though high concentrations/long exposure could affect nuclear staining.

Conclusions:

  • PTA is a valuable micro-CT contrast agent for nerve imaging, particularly for visualizing the perineurium.
  • PTA demonstrates minimal impact on histological integrity, supporting its use in multimodal studies.
  • This technique aids in understanding nerve organization for improved therapeutic strategies.