研究目的
Investigating the utility of ICG topically applied to the colon surface in the imaging of proliferative lesions of the colon.
研究成果
Fluorescence contrast-enhanced imaging with ICG enema enhances detection of colon proliferative lesions, attributed to tight junction disruption promoting endocytosis and ICG uptake in tumor cells.
研究不足
The low penetrability of ICG limits detection to surface epithelia, and longer retention times are impractical for clinical use.
1:Experimental Design and Method Selection:
The study used a rat model of azoxymethane-induced colon carcinogenesis to evaluate ICG uptake via enema administration, employing live-cell imaging, in vivo endoscopy, ex vivo microscopy, and immunofluorescence microscopy to assess cellular and tissue responses.
2:Sample Selection and Data Sources:
Female F344 rats were used, with colon tumor cell line RCN-9 for in vitro experiments. Samples included colon tissues from rats with induced carcinogenesis.
3:List of Experimental Equipment and Materials:
Fluorescence imaging systems (OV100, IV100, VS120-FL), endoscopic system with fiberscope (BF-XP60), ICG (Diagnogreen), organelle markers, inhibitors like sulfobromophthalein, and antibodies for immunofluorescence.
4:Experimental Procedures and Operational Workflow:
Rats were administered ICG enema, retained for 30 minutes, followed by imaging. Cultured cells were incubated with ICG under various conditions (e.g., temperature, inhibitors) and imaged. Tissues were excised, stained, and analyzed.
5:Data Analysis Methods:
Mean fluorescence intensity was measured using software, and immunofluorescence was analyzed to assess protein expression and localization.
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OV100
OV100
Olympus Corporation
Fluorescence reflectance imaging system for microscopic imaging
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IV100
IV100
Olympus Corporation
Multi-wavelength laser scanning microscope for high-power imaging
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VS120-FL
VS120-FL
Olympus Corporation
Fluorescence virtual microscopy system for scanning live cells and tissue sections
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BF-XP60
BF-XP60
Olympus Medical Corporation
Flexible endoscope fiberscope for in vivo endoscopic imaging
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Hoechst 33342
Hoechst 33342
Thermo Fisher Scientific
Nuclear staining dye for live-cell imaging
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Sulfobromophthalein
BSP
Sigma-Aldrich
Membrane transporter inhibitor used in cellular uptake experiments
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WGA conjugated with Alexa Fluor 555
WGA Alexa Fluor 555
Thermo Fisher Scientific
Wheat germ agglutinin conjugated for mucus imaging
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MC285SPD-L0B0
MC285SPD-L0B0
Texas Instruments
Sensitive electron-multiplying charge-coupled device camera for detecting fluorescence
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Diagnogreen
Diagnogreen
Daiichi-Sankyo
Pharmaceutical grade indocyanine green used as a fluorescent contrast agent
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TL488
TL488
Vector Laboratories
Lycopersicon esculentum (tomato) lectin conjugated with DyLight 488 for blood vessel visualization
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