高HCY培养内皮细胞表面形态的AFM观察

THE ENDOTHELIAL CELL SURFACE CHANGE IN DIFFERENTCONCENTRATIONS OF HOMOCYSTEINE BY ATOMIC FORCE MICROSCOPE

目的：探讨一种新的损伤小、分辨率高的观察内皮细胞表面形态的方法。方法：取新生胎儿脐带静脉血管内皮细胞，培养２４～４８小时后，分为高ＨＣＹ实验组和正常对照组，采用ＡＦＭ对用戊二醛固定与未固定的内皮细胞进行观察，并与倒置显微镜、电子显微镜进行比较。结果：（１）固定后的内皮细胞ＡＦＭ扫描图象见细胞表面颗粒界限分明，排列规则，平均尺度为３００～４００ｎｍ，平整度略好于培养条件下观察的内皮细胞图象；（２）固定的高ＨＣＹ实验组内皮细胞的ＡＦＭ扫描图象见细胞表面有非常多的突起，颗粒规则性排列的特征消失，颗粒间界限模糊，表面粗糙，呈虫蚀状变化。（３）倒置显微镜观察未见两组间差异，电子显微镜观察也无法分辨差异。结论：ＡＦＭ是一种观察内皮细胞表面形态的较好方法，损伤小。

A new surface scan method, which has less injury and higher resolution, is applied to observe endothelial cell(EC). After cultured 24-48 hours, the VEC is divided into 2 groups:one is treated with higher homocysteine, the other is control. AFM is used to observe the cell which is treated with or without glutaraldehyde(HCY), the image is compared with its optical microscope or EM. 1) After cultured without HCY and fixed with glutaraldehyde, the EC surface has clear cut and regular arranged granular structure, the size ranging from 300-400nm; 2) After cultured with HCY and fixed with glutaraldehyde, the cell's surface become rough; and bulged granular-flatted the boundary, and erode the surface; 3) LM couldn't distinguish the difference, EM couldn't either. AFM not only has a higher resolution than LM, EM, but also has less prerequisite, which is essential for EM.