Ex-vivo study of high-magnification chromoendoscopy in the gastrointestinal tract to determine the optimal staining conditions for endocytoscopy.
OBJECTIVE
Endocytoscopy allows the observation of living cells in the gastrointestinal tract. Consistently clear views are essential for clinical application of the technique, but these are not always obtained. The aim of this study was to determine an appropriate staining regimen for endocytoscopy.


METHODS
This was an ex-vivo animal study in which we stained freshly resected porcine esophagus, stomach, and colon with different concentrations of three dyes (1%, 0.5%, and 0.25% crystal violet; 5%, 2.5%, and 1% methylene blue; and 1%, 0.5%, and 0.25% toluidine blue) and assessed them after different exposure times (10 seconds, 30 seconds, 60 seconds, and 90 seconds). The images obtained were evaluated according to the staining status of the cytoplasm and the nucleus, and the contrast between the cytoplasm and the nuclei, and the optimal staining conditions for each organ were determined. Additionally, freshly resected human esophagus, stomach, and colon tissues were tested under the dye/exposure conditions that were found to be the most appropriate in the animal study.


RESULTS
After intensive mucus removal, high-quality images were obtained using methylene blue and toluidine blue. The optimum conditions for endocytoscopic observation were obtained after staining with 1% methylene blue in the esophagus and with 0.25% toluidine blue in the stomach and the colon, after 60 seconds of exposure to the dye. This was confirmed in the human specimens.


CONCLUSIONS
This study provides important information on appropriate staining conditions for endocytoscopy. Further ex-vivo and in-vivo studies are necessary before this technique comes into standard use, however.
Kodashima
S
S
Department of Gastroenterology, The University of Tokyo, Tokyo, Japan.
Fujishiro
M
M
Takubo
K
K
Kammori
M
M
Nomura
S
S
Kakushima
N
N
Muraki
Y
Y
Tateishi
A
A
Kaminishi
M
M
Omata
M
M
D000818:Animals; D003106:Colon / Q000033:anatomy & histology*; D004396:Coloring Agents / Q000008:administration & dosage; D016099:Endoscopy, Gastrointestinal / Q000379:methods*; D004947:Esophagus / Q000033:anatomy & histology*; D005240:Feasibility Studies; D005840:Gentian Violet / Q000008:administration & dosage; D006801:Humans; D008751:Methylene Blue / Q000008:administration & dosage; D013194:Staining and Labeling / Q000379:methods*; D013270:Stomach / Q000033:anatomy & histology*; D013552:Swine; D046509:Tissue Culture Techniques; D014048:Tolonium Chloride / Q000008:administration & dosage
D004396:Coloring Agents; D014048:Tolonium Chloride; D005840:Gentian Violet; D008751:Methylene Blue