扫描探针显微术用于B淋巴瘤细胞表面分子数量化与可视化研究

Visualization and quantitation of CD20 molecues on human B-cell lymphoma by scanning probe microscopy (SPM)

本研究试图采用原子力显微术(AFM)和近场光学显微术(NSOM)对B淋巴瘤细胞的表面形态和超微结构进行了纳米级高分辨的成像研究,获得Rituximab抗体处理前后不同状态下的高清晰、对比度良好的NSOM和AFM的B淋巴瘤细胞图像,建立超高分辨率显微镜观测B淋巴瘤细胞膜分子表面分布的方法。采用AFM和NSOM,对B淋巴瘤细胞的表面形态及光学性质进行了观测。结果显示建立了一种观测B淋巴瘤细胞超微细胞表面结构的方法,结合NSOM对Rituximab抗体处理前后CD20膜蛋白分子在细胞膜表面位置进行高分辨率的观测。AFM的结果表明Rituximab抗体处理以后B淋巴瘤细胞细胞膜的颗粒度明显增加,NSOM结果发现在Rituximab处理B淋巴瘤细胞前后,CD20膜蛋白分子仍然散在分布在细胞膜表面。

AFM （atomic force microscopy） and NSOM （near field scanning microscopy） have extremely high resolution and many advantages in exploring the biology. In this work, AFM and NSOM were used to investigate the morphology and ultrastructure of human B-cell lymphoma before and after treatment with CD20 antibody Rituximab, we used AFM combined with NSOM to optically investigate CD20 molecules on human Bcell lymphoma. The location, distribution and variation of CD20 molecules were imaged by a high-resolve NSOM. The results showed that a method that could be used to observe the ultrastructure of surface molecules on B cell lymphoma was established and the distribution of membrane-bound CD20 molecules was observed by the two types of high resolution microscopies. After treatment with Rituximab, the lymphoma cell surface has small granules, indicating abundant structure infomation inside the cells, and NSOM found that CD20 molecules remained scat tered on the cell surface.