Li Hung-Wing, McCloskey Michael, He Yan, Yeung Edward S
Ames Laboratory-USDOE and Department of Chemistry, Iowa State University, Ames, IA 50011, USA.
Anal Bioanal Chem. 2007 Jan;387(1):63-9. doi: 10.1007/s00216-006-0403-8. Epub 2006 Apr 22.
We demonstrate the capability of differential interference contrast (DIC) microscopy as a simple and useful tool for studying cellular events without fluorescence labeling. By coupling an advanced DIC microscope to a computer-controlled motorized vertical stage and a high-speed, high-resolution CCD camera, real-time three-dimensional monitoring is possible in a high-throughput manner. The performance among three modes of microscopy, bright-field, dark-field and DIC, in terms of horizontal resolving power and vertical sectioning was investigated. As a model, exocytosis of rat peritoneal mast cells was recorded on the subsecond time scale. Three-dimensional tracking of granules during degranulation was achieved and granule-granule fusion before plasma membrane fusion was recorded.
我们证明了微分干涉差（DIC）显微镜作为一种无需荧光标记即可研究细胞事件的简单且有用工具的能力。通过将先进的DIC显微镜与计算机控制的电动垂直载物台和高速、高分辨率的电荷耦合器件（CCD）相机相结合，可以以高通量方式进行实时三维监测。研究了明场、暗场和DIC这三种显微镜模式在水平分辨率和垂直切片方面的性能。作为一个模型，在亚秒时间尺度上记录了大鼠腹膜肥大细胞的胞吐作用。实现了脱颗粒过程中颗粒的三维跟踪，并记录了质膜融合前颗粒与颗粒的融合。
