Sun Wei, Xiao Lehui, Fang Ning
Ames Laboratory, U.S. Department of Energy and Department of Chemistry, Iowa State University, Ames, IA, USA.
Methods Mol Biol. 2013;931:169-86. doi: 10.1007/978-1-62703-056-4_10.
Optical microscopy is a simple yet robust strategy to study live cellular processes. By changing the wavelength of the illumination light, different non-fluorescent nanoparticle probes can be identified and tracked dynamically inside crowded living cells with either differential interference contrast (DIC) microscopy or planar illumination microscopy (PIM). The translational and rotational dynamics of anisotropic nanoparticles can be readily extracted via the modified DIC microscope and the home-built PIM. In this protocol, the optimization procedures for DIC microscopy and PIM imaging are explained, and the sample preparation procedures to image non-fluorescent nanoparticles in living cells are described.
光学显微镜是一种研究活细胞过程的简单而可靠的策略。通过改变照明光的波长，可以使用微分干涉对比（DIC）显微镜或平面照明显微镜（PIM）在拥挤的活细胞内动态识别和跟踪不同的非荧光纳米颗粒探针。各向异性纳米颗粒的平移和旋转动力学可以通过改进的DIC显微镜和自制的PIM轻松提取。在本方案中，解释了DIC显微镜和PIM成像的优化程序，并描述了对活细胞中的非荧光纳米颗粒进行成像的样品制备程序。
