研究目的
To develop a simple optical method for direct noninvasive CTC detection using confocal microscopes, enabling continuous, real-time, and long-duration detection of CTCs in animal models.
研究成果
Confocal microscopy can be used as a simple and reliable method to monitor CTCs in vivo for cancer research, enabling detection of rare CTCs at the early stage of tumor development and distinguishing CTC clusters from single CTCs.
研究不足
The confocal setup might miss out-of-focus CTC signals, resulting in lower sensitivity for CTCs in the whole blood vessel. The method requires fluorescent labeling of CTCs, which may not be applicable for all cancer types.
1：Experimental Design and Method Selection:
The study utilized confocal microscopy for CTC detection, focusing on line scanning across blood vessels to excite and detect fluorescently-labeled CTCs.
2：Sample Selection and Data Sources:
Human hepatocellular carcinoma cell line (HCCLM3) and prostate cancer cell line (PC3) were used, with cells genetically labeled with EGFP for fluorescence detection.
3：List of Experimental Equipment and Materials:
Confocal microscope system based on an inverted microscope (IX73, Olympus), lasers at 488 and 532 nm, PMT detectors, DiI for fluorescence labeling.
4：Experimental Procedures and Operational Workflow:
Mice were anesthetized, tumor cells were injected or implanted, and confocal line scanning was performed on blood vessels in the ear for CTC detection.
5：Data Analysis Methods:
Fluorescence signals were analyzed to distinguish between single CTCs and CTC clusters based on signal width and peak characteristics.
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