三维培养结肠癌细胞的微流控芯片荧光成像研究

Investigation of fluorescence imaging in microfluidic chip for three-dimensional cultivation of colon cancer cells

改良了一种微流控芯片,可用于对结肠癌细胞进行三维培养并实现实时荧光成像。在结肠癌细胞内植入内源性的红色荧光蛋白,使用激光共聚焦显微镜对芯片中三维培养的细胞进行成像。通过细胞内部红色荧光蛋白的表达,可以观测到细胞的生长状态,实现对细胞的实时监测和高分辨率荧光成像。同时,通过免疫荧光染色来表征反映细胞活性的特征蛋白,其荧光强度和蛋白表达呈正相关。研究结果提示,细胞活性相关蛋白的表达受到微环境的影响,其在芯片三维培养中的活性强于二维培养,表明芯片内环境更加接近真实的人体微环境。该方法为进一步探究肿瘤细胞转移机制及相关药物的筛选研究提供了一种新的技术手段及实验平台。

This article presents an improved method for the three-dimensional cultivation of colon cancer cells and the realization of real-time fluorescence imaging using a microfluidic chip.By incorporating endogenous red fluorescent protein into the colon cancer cells,we employed laser confocal microscopy to visualize the cells cultivated within the chip in three dimensions.The expression of intracellular red fluorescent protein allowed for the observation of cell growth status,facilitating real-time monitoring and high-resolution fluorescence imaging.Moreover,immunofluorescence staining was employed to characterize feature proteins indicative of cellular activity,with their fluorescence intensity demonstrating a positive correlation with protein expression.The research findings indicate that the expression of activity-related proteins is influenced by the microenvironment,exhibiting stronger activity in the three-dimensional cultivation on the chip compared to the two-dimensional approach.This suggests that the microenvironment within the chip more closely resembles the actual microenvironment within the human body.This method provides a novel technical approach and experimental platform for further exploration of tumor metastasis mechanisms and the screening of related drugs.