Emiliani Valentina, Cojoc Dan, Ferrari Enrico, Garbin Valeria, Durieux Christiane, Coppey-Moisan Maite, Di Fabrizio Enzo
Opt Express. 2005 Mar 7;13(5):1395-405. doi: 10.1364/opex.13.001395.
A new method to perform simultaneously three dimensional optical sectioning and optical manipulation is presented. The system combines a multi trap optical tweezers with a video microscope to enable axial scanning of living cells while maintaining the trapping configuration at a fixed position. This is achieved compensating the axial movement of the objective by shaping the wave front of the trapping beam with properly diffractive optical elements displayed on a computer controlled spatial light modulator. Our method has been validated in three different experimental configurations. In the first, we decouple the position of a trapping plane from the axial movements of the objective and perform optical sectioning of a circle of beads kept on a fixed plane. In a second experiment, we extend the method to living cell microscopy by showing that mechanical constraints can be applied on the dorsal surface of a cell whilst performing its fluorescence optical sectioning. In the third experiment, we trapped beads in a three dimensional geometry and perform, always through the same objective, an axial scan of the volume delimited by the beads.
提出了一种同时进行三维光学切片和光学操控的新方法。该系统将多阱光镊与视频显微镜相结合，能够在保持捕获配置固定在一个位置的同时对活细胞进行轴向扫描。这是通过在计算机控制的空间光调制器上显示的适当衍射光学元件对捕获光束的波前进行整形来补偿物镜的轴向移动而实现的。我们的方法已在三种不同的实验配置中得到验证。在第一种配置中，我们将捕获平面的位置与物镜的轴向移动解耦，并对固定平面上的一圈珠子进行光学切片。在第二个实验中，我们通过表明在对细胞进行荧光光学切片时可以对细胞的背表面施加机械约束，将该方法扩展到活细胞显微镜检查。在第三个实验中，我们将珠子捕获在三维几何结构中，并始终通过同一个物镜对由珠子界定的体积进行轴向扫描。
