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Confocal Fluorescence Microscopy

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Probe-based Confocal Laser Endomicroscopy of the Urinary Tract: The Technique

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Technical feasibility of visualizing myenteric plexus using confocal laser endomicroscopy.

Masakuni Kobayashi¹, Kazuki Sumiyama², Naoki Shimojima^3,4

¹Department of Endoscopy and Division of Gastroenterology and Hepatology, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan.

Journal of Gastroenterology and Hepatology

|February 1, 2017

Summary

This study shows that cresyl violet (CV) can be safely used with probe-based confocal laser endomicroscopy (pCLE) for visualizing the myenteric plexus (MP) from the serosal side in human surgical specimens.

Keywords:

clinical pediatric gastroenterology endoscopy: colon pediatric disorders

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Area of Science:

Gastroenterology
Surgical Endoscopy
Neuroscience

Background:

Previous myenteric plexus (MP) visualization using confocal laser endomicroscopy (CLE) relied on neural probes without established clinical safety.
Visualization was previously performed from the submucosal side.

Purpose of the Study:

To evaluate the technical feasibility of visualizing the MP from the serosal side using probe-based CLE (pCLE) with cresyl violet (CV).
To assess the safety and efficacy of CV for MP visualization in human surgical specimens.

Main Methods:

Explored CV dye affinity for MP in a transgenic mouse model.
Assessed CV-assisted pCLE visualization of MP in human surgical specimens from the serosal side.
Evaluated visualization rate, sensitivity, and specificity against pathological standards.

Main Results:

Confirmed CV dye affinity for MP in all models.
MP visualized in 85.7% of human samples via CV-assisted pCLE with high sensitivity (92.3%) and specificity (100%).
pCLE identified ladder-like nerve structures and ganglia, with mean nerve strand widths of 54.3 μm (max) and 19.7 μm (min).

Conclusions:

Demonstrated the technical feasibility of real-time MP visualization using CV-assisted pCLE from the serosal side.
CV is a viable and safe option for intraoperative MP assessment.