In vivo histologic imaging of the muscularis propria and myenteric neurons with probe-based confocal laser endomicroscopy in porcine models (with videos)

Ohya, Tomohiko R.; Sumiyama, Kazuki; Takahashi‐Fujigasaki, Junko; Dobashi, Akira; Saito, Shôichi; Tajiri, Hisao

doi:10.1016/j.gie.2011.09.045

Cited by 30 publications

(26 citation statements)

References 27 publications

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“…This is consistent with a previous study that demonstrated visualization by NeuroTrace of the cytoplasm (and not nuclei) in neuron-like cells. 25 We also showed that NeuroTrace stains strongly both the cytoplasm and nuclei of glial cells, which provide support and protection for neurons and also serve as neuronal cell precursors in vivo. [26][27][28] Furthermore, our immunostaining studies for NGF expression demonstrated for the first time a strong expression of NGF in the same cells that exhibit staining with NeuroTrace indicating that GENS cells synthesize NGF.…”

Section: Discussionmentioning

confidence: 56%

“…A recent study used a 2.7-mm-diameter pCLE probe to visualize the ENS, but this required an invasive procedure-endoscopic resection of the submucosa. 25 Our study used a novel, needle-based CLE (nCLE) probe introduced into gastric submucosa and muscularis propria (under EUS guidance) and local administration of NeuroTrace through a fine needle. The field of view of nCLE probe is larger than pCLE probe (325 μm vs 240 μm), which likely has some advantages in this particular application also.…”

Section: Discussionmentioning

confidence: 99%

“…The application of NeuroTrace for the human use is currently limited because the human safety of NeuroTrace injection is unknown, but some clinically approved fluorescence Nissl stains-fluorescein and cresyl violet-are in a testing phase. 25 Another potential promising technique for visualizing enteric ganglia in the gastrointestinal tract is optical coherence microscopy; however, it lacks the high resolution of CLE. 29 This current study was an early pilot study to determine if in vivo imaging of GENS using needle-based CLE was feasible.…”

Section: Discussionmentioning

confidence: 99%

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In vivo imaging of porcine gastric enteric nervous system using confocal laser endomicroscopy &molecular neuronal probe

Samarasena

Ahluwalia

Shinoura

et al. 2016

J of Gastro and Hepatol

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Section: Discussionmentioning

confidence: 56%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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In vivo imaging of porcine gastric enteric nervous system using confocal laser endomicroscopy &molecular neuronal probe

Samarasena

Ahluwalia

Shinoura

et al. 2016

J of Gastro and Hepatol

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“…The limitations of these 2 techniques are the small sample volume of MP and the fragmented samples that can impair proper tissue orientation and impact diagnostic yield. The use of confocal laser endomicroscopy for optical histologic imaging is promising, but depth of imaging may not be sufficient in the stomach and the applicability of dye-based methods in humans is unclear 18,19,20,21 .…”

Section: Discussionmentioning

confidence: 99%

Innovative gastric endoscopic muscle biopsy to identify all cell types, including myenteric neurons and interstitial cells of Cajal in patients with idiopathic gastroparesis: a feasibility study (with video)

Rajan

Gostout

Song

et al. 2016

Gastrointestinal Endoscopy

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Background and Aims The pathophysiology of some gastrointestinal neuromuscular diseases (GINMD) remains largely unknown. This is in part due to the inability to obtain ample deep gastric wall biopsies that include the intermuscular layer of the muscularis propria (MP) to evaluate the enteric nervous system, interstitial cells of Cajal (ICC) and related cells. We report on a novel technique for gastric endoscopic muscle biopsy (gEMB). Methods Patients with idiopathic gastroparesis were prospectively enrolled in a feasibility study using a novel “no hole” gEMB. Main outcome measures were technical success, adverse events and histological confirmation of the intermuscular layer, including myenteric neurons and ICC. The gEMB was a double resection clip–assist technique. A site was identified on the anterior wall of the gastric body as recommended by the International Working Group on histological techniques. Endoscopic mucosal resection was performed to unroof and expose the underlying MP. The exposed MP was then retracted into the cap of an over-the-scope clip. The clip was deployed and the pseudopolyp of MP created was resected. This resulted in a “nohole” gEMB. Results Three patients with idiopathic gastroparesis underwent gEMB. Patients had severe delayed gastric emptying with a mean of 49 ± 16.8% of retained gastric contents at 4 hours. They had no history of gastric or small-bowel surgery, and did not use steroids or other immunosuppressive drugs. The gEMB procedure was successfully performed with no procedural adverse events. Postprocedure abdominal pain was controlled with nonsteroidal anti-inflammatory agents and opioid analgesics. Mean length of resected MP was 10.3 ± 1.5 mm. Mean procedure time was 25.7 ± 6 minutes. Tissue samples on hematoxylin and eosin (H&E) stain confirmed presence of both inner circular and outer longitudinal muscle as well as the intermuscular layer. H&E stain showed reduced myenteric ganglia in 1 patient. In 2 patients, specialized immunohistochemistry was performed, which showed marked decrease in myenteric neurons as delineated by an antibody to Protein Gene Product (PGP) 9.5 and severe decrease in ICC across the muscle layers. At 1 month follow-up, upper endoscopy showed a well-healed scar in 2 patients and minimal ulceration with retained clip in 1 patient. CT abdomen confirmed the integrity of the gastric wall in all patients. Due to lack of an immune infiltrate in the resected samples, patients were not considered suitable for immunosuppressive or steroid therapy. Conclusions gEMB is feasible and easy to perform, with acquisition of tissue close to surgical samples to identify myenteric ganglia, ICC and multiple cell types. The ability to perform gEMB represents a paradigm shift in endoscopic tissue diagnosis of gastric neuromuscular pathologies.

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“…As the pCLE technique uses light in the VIS spectral region no side effects should be occur. Due to the confocal technique, tissue microarchitecture and cellular morphology in a tissue depth of up to 60 μm could be visualized at a histomorphological level [16,17]. PCLE enables in vivo microscopy of tissues by mean of flexible miniaturized fiberoptic probes of 1.5 to 2.6 mm outer diameter inserted through the working channel of a standard endoscope.…”

Section: Introductionmentioning

confidence: 99%

Probe‐based confocal laser endomicroscopy (pCLE) – a new imaging technique for in situ localization of spermatozoa

Trottmann

Stepp

Sroka

et al. 2014

Journal of Biophotonics

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In azoospermic patients, spermatozoa are routinely obtained by testicular sperm extraction (TESE). However, success rates of this technique are moderate, because the site of excision of testicular tissue is determined arbitrarily. Therefore the aim of this study was to establish probe‐based laser endomicroscopy (pCLE) a noval biomedical imaging technique, which provides the opportunity of non‐invasive, real‐time visualisation of tissue at histological resolution. Using pCLE we clearly visualized longitudinal and horizontal views of the tubuli seminiferi contorti and localized vital spermatozoa. Obtained images and real‐time videos were subsequently compared with confocal laser scanning microscopy (CLSM) of spermatozoa and tissues, respectively.

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In vivo histologic imaging of the muscularis propria and myenteric neurons with probe-based confocal laser endomicroscopy in porcine models (with videos)

Cited by 30 publications

References 27 publications

In vivo imaging of porcine gastric enteric nervous system using confocal laser endomicroscopy &molecular neuronal probe

In vivo imaging of porcine gastric enteric nervous system using confocal laser endomicroscopy &molecular neuronal probe

Innovative gastric endoscopic muscle biopsy to identify all cell types, including myenteric neurons and interstitial cells of Cajal in patients with idiopathic gastroparesis: a feasibility study (with video)

Probe‐based confocal laser endomicroscopy (pCLE) – a new imaging technique for in situ localization of spermatozoa

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