Aix-Marseille Université, Ecole Centrale Marseille, CNRS UMR 7249, Institut Fresnel, Marseille, France.
J Biomed Opt. 2012 Jul;17(7):076004. doi: 10.1117/1.JBO.17.7.076004.
We present a quadriwave lateral shearing interferometer used as a wavefront sensor and mounted on a commercial non-modified transmission white-light microscope as a quantitative phase imaging technique. The setup is designed to simultaneously make measurements with both quantitative transmission phase and fluorescence modes: phase enables enhanced contrasted visualization of the cell structure including intracellular organelles, while fluorescence allows a complete and precise identification of each component. After the characterization of the phase measurement reliability and sensitivity on calibrated samples, we use these two imaging modes to measure the characteristic optical path difference between subcellular elements (mitochondria, actin fibers, and vesicles) and cell medium, and demonstrate that phase-only information should be sufficient to identify some organelles without any labeling, like lysosomes. Proof of principle results show that the technique could be used either as a qualitative tool for the control of cells before an experiment, or for quantitative studies on morphology, behavior, and dynamics of cells or cellular components.
我们提出了一种用于波前传感器的四波横向剪切干涉仪，并将其安装在商业非改装的透射白光显微镜上，作为一种定量相位成像技术。该设置旨在同时以定量透射相位和荧光两种模式进行测量：相位能够增强细胞结构的对比度可视化，包括细胞内细胞器，而荧光则允许对每个组件进行完整和精确的识别。在对校准样品的相位测量可靠性和灵敏度进行了表征之后，我们使用这两种成像模式来测量亚细胞元件（线粒体、肌动蛋白纤维和囊泡）与细胞介质之间的特征光程差，并证明仅相位信息就足以识别一些无需任何标记的细胞器，如溶酶体。原理验证结果表明，该技术既可以用作实验前细胞控制的定性工具，也可以用于细胞或细胞成分的形态、行为和动力学的定量研究。
