Creath Katherine
Optineering, Tucson, AZ USA 85719, and College of Optical Sciences, The University of Arizona, Tucson, AZ USA 85721.
Proc SPIE Int Soc Opt Eng. 2010 Aug 2;7782:77820B-. doi: 10.1117/12.864275.
This paper presents images and data of live biological samples taken with a novel Linnik interference microscope. The specially designed optical system enables instantaneous and 3D video measurements of dynamic motions within and among live cells without the need for contrast agents. This "label-free", vibration insensitive imaging system enables measurement of biological objects in reflection using harmless light levels with current magnifications of 10X (NA 0.3) and 20X (NA 0.5) and wavelengths of 660 nm and 785 nm over fields of view from several hundred microns up to a millimeter. At the core of the instrument is a phase-measurement camera (PMC) enabling simultaneous measurement of multiple interference patterns utilizing a pixelated phase mask taking advantage of the polarization properties of light. Utilizing this technology enables the creation of phase image movies in real time at video rates so that dynamic motions and volumetric changes can be tracked. Objects are placed on a reflective surface in liquid under a coverslip. Phase values are converted to optical thickness data enabling volumetric, motion and morphological studies. Data from a number of different mud puddle organisms such as paramecium, flagellates and rotifers will be presented, as will measurements of flying ant wings and cultures of human breast cancer cells. These data highlight examples of monitoring different biological processes and motions. The live presentation features 4D phase movies of these examples.
本文展示了用新型 Linnik 干涉显微镜拍摄的活生物样本的图像和数据。经过特殊设计的光学系统能够对活细胞内部和之间的动态运动进行即时三维视频测量,而无需使用造影剂。这种“无标记”、对振动不敏感的成像系统能够在无害的光水平下,以当前 10 倍(数值孔径 0.3)和 20 倍(数值孔径 0.5)的放大倍数以及 660 纳米和 785 纳米的波长,在从几百微米到一毫米的视场范围内对生物物体进行反射测量。该仪器的核心是一台相位测量相机(PMC),它利用像素化相位掩膜并借助光的偏振特性,能够同时测量多个干涉图样。利用这项技术可以以视频速率实时创建相位图像电影,从而能够跟踪动态运动和体积变化。物体放置在盖玻片下液体中的反射表面上。相位值被转换为光学厚度数据,可用于体积、运动和形态学研究。将展示来自多种不同泥坑生物(如草履虫、鞭毛虫和轮虫)的数据,以及飞蚁翅膀的测量数据和人类乳腺癌细胞培养的数据。这些数据突出了监测不同生物过程和运动的实例。现场展示将呈现这些实例的 4D 相位电影。