原子力显微镜新应用：单细胞诊断肝癌

New application of atomic force microscopy., diagnosis of hepatocellular carcinoma by one cell

目的运用原子力显微镜（AFM）分析正常肝细胞、肝硬化细胞、肝癌细胞膜表面微细结构；绘出细胞膜表面蛋白变化图，并以此图作为诊断肝癌的标准。方法选取正常肝细胞、肝硬化细胞、肝癌细胞各40组，利用AFM对标本扫描，将细胞膜表面扫描图用分析软件进行分析。结果正常肝细胞、肝硬化细胞、肝癌细胞的细胞膜表面结构存在明显的差异，从正常肝细胞到肝硬化细胞再到肝癌细胞是一个渐进的过程。每个细胞膜表面的蛋白微细结构分布并不均匀，从膜表面蛋白分析数据中可看到它们存在差异。结论依据细胞膜表面分析数据做出三组图形，我们可以以此为肝癌的诊断标准，只要获得一个细胞，即可做出肝癌的诊断。根据细胞膜表面蛋白分布的不同，将细胞膜表面分为功能活跃区和功能不活跃区。

Objective To use atomic force microscopy to analyze the surface microstructure of normal liver cells, cirrhosis cells and liver cancer cells to draw the protein map of changes in the surface membrane, make the diagnosis standards of hepatocellular carcinoma and diagnose hepatocellular carcinoma by just 1 liver cell. Methods We collected normal liver cells, cirrhotic cells and hepatoma cells（40 examples for each group）, fixed them with 2% formalin, and made them into print slices and then dried them in clean space. A number of 10 cells of each sample was scanned with AFM using tapping mode in the range of 30 um to 500 nm and at the rate of 0.5-2 Hz. The surface of cell membrane was analyzed with analysis software. Results In the progress from normal hepatocytes to cirrhotic cells and then hepatoma cells, the fine structure of cell membrane surface changed accordingly and obviously, and the membrane proteins increased gradually, with apophysis increasing gradually shown in the AFM. In normal liver cells, cirrhosis, liver cancer cells in the membrane surface structure, there was an obvious difference. Cirrhosis of the liver cells from normal cells and then hepatoma cells was a gradual process. Micro membrane proteins were not well-distributed in the cell membrane surface. Analysis of data from the surface protein showed their differences. Conclusion Based on the cell membrane surface graph, we can use this standard in the diagnosis of hepatocellular carcinoma. According to the distribution of different cell surface protein, the membrane surface is divided into functional active areas and functional inactive area.