Colored point spread function engineering for parallel confocal microscopy.
Colored point spread function engineering for parallel confocal microscopy.
复制标题
用于平行共焦显微镜的彩色点扩散函数工程。
作者：
A. Jesacher;S. Bernet;M. Ritsch
利用空间光调制器(SLM)对两者的颜色选择性，在一个波长上定制激发光束，并在红移的荧光波长上多路复用图像，可以使共焦显微镜并行化，即同时检测样品的轴向堆叠(Z堆叠)。为此，两个衍射图案，一个控制激发光，另一个操纵发射光，在SLM的同一区域内组合，充当纯相位调制器。最近展示的一种技术允许人们使用SLM的扩展相移能力来组合具有高衍射效率和低串扰的图案，该相移能力在各个波长覆盖2π的倍数。在第一个演示中，我们比较了标准共焦成像和在两个独立样本平面上的同时图像采集，这显示了可比较的结果。
Using the color selectivity of a spatial light modulator (SLM) for both, tailoring the excitation beam at one wavelength, and multiplexing the image at the red-shifted fluorescence wavelength, it is possible to parallelize confocal microscopy, i.e. to simultaneously detect an axial stack (z-stack) of a sample. For this purpose, two diffractive patterns, one steering the excitation light, and the other manipulating the emission light, are combined within the same area of the SLM, which acts as a pure phase modulator. A recently demonstrated technique allows one to combine the patterns with high diffraction efficiency and low crosstalk, using the extended phase shifting capability of the SLM, which covers multiples of 2π at the respective wavelengths. For a first demonstration we compare standard confocal imaging with simultaneous image acquisition in two separate sample planes, which shows comparable results.