暗场聚光镜-水浸物镜联在细胞成像上的初步应用

Construction of awater immersion objective-darkfield condenser combination optical system and its initial application in cell image

传统的细胞成像方法在准备工作中需要对细胞进行多步操作,对细胞的实时成像较难以反映真实生长环境中的活细胞.本实验引入了一种高数值孔径的油浸暗场聚光镜与无盖玻片水浸物镜联用来进行高分辨率实时成像的方法.为了尽可能地减少细胞操作步骤,本实验采用水浸物镜直接伸入到培养皿中的PBS(磷酸缓冲生理盐水)或培养基中的方法取代传统的细胞爬片与细胞滴片操作,来得到贴壁培养与悬浮培养细胞的实时暗场图像.用这一系统观察PBS里的贴壁细胞能得到较好的暗场成像效果,观察培养基中悬浮细胞可得到细胞图像,但视野过亮.这一组合的光学系统简便易行,成本低廉,可广泛用于细胞相关的科研工作中.

The traditional method of cell imaging is too much operation on the cells in the preparation work.It is not suitable for real-time observation of living cells.This experiment introduces a method of combining a large numerical aperture oil immersed dark-field condenser and a coverslip-less water immersion objective to high resolution real-time cell imaging.To minimize cell operation in the preparations for the real-time imaging,this experiment by using water immersion lens inserted directly into the Petri dish in PBS(phosphate Physiological Buffered Saline)or culture medium to replace the traditional cell smear and cell drop operation,to obtain adherent and suspension cultured cells real-time dark field image culture.Observation of adherent cells in PBS can get better effect in this dark field imaging system;observation of suspension cell in culture medium can be obtained,but the dark field effect.This optical system is simple and inexpensive,and can be widely used in the research work related to cells.