应用原子力显微镜技术对人膀胱癌细胞株T24活细胞的成像研究

Using the AFM to observe and study the ultrastructure of the living T24 cells of the human bladder cancer

目的实现应用原子力显微镜（AFM）技术对人膀胱癌细胞株124活细胞的成像，在原子级或纳米级水平上观测T24活细胞的超微结构。方法在生理条件下，使用AFM直接观测培养中的人膀胱癌T24细胞，对其超微结构动态成像。结果获得T24活细胞原子级或纳米级分辨率下的实时扫描图像，细胞胞膜和细胞骨架成像清晰。结论AFM技术可以用于对膀胱癌T24活细胞超微结构的成像研究，所获得的图像可反映其在生活状态下的动态变化。

Objective By using atomic force microscopy （AFM） to observe and study the ultrastructure of the living T24 cells of the human bladder cancer. Simultaneously, according to these images the dynamic change of the cancerous epithelial cells of the bladder mucosa was revealed at the atomic or molecular level. Methods The T24 cells of the human bladder cancer were selected and cultured in a 95% air and 5% CO2 incubator at 37℃. They were grown in RPMI1640 medium （pH 7.4） containing 10% fetal calf serum （FCS）. The dishes （35mm × 35mm） which had been used to culture the cells, were treated in 5% Gelatin, so the viscosity of the dishes could be increased, and the scanning procedure could be completed successfully. On top of, the ultrastructure of these living cells had been observed using the AFM （NanoⅢa, Digital Instrument Company） on physiological conditions at the constant temperature controlled by a thermostat （ Western Germany Opton TRZ3700）. Results The AFM images of the living T24 cells had been attained, and the images of the membrane and the cytoskeleton were very clear. Conclusions The AFM was a novel method to observe and analysis the ultrastructure of the living cells of the human bladder cancer directly and real-timely. This experimental model would play an important role for elucidating the cancerous mechanism of the bladder normal cells at the atomic or nanometer level.