基于数字微镜装置的结构光照明层析成像

摘要

在结构光显微系统中使用可编程数字微镜装置代替压电驱动的光栅,产生稳定可靠的结构光照明.为避免数字微镜装置本身的像素化结构对成像的影响,系统中引入一个低通的空间光调制器,以滤除经过数字微镜装置后产生的高级(±2nd及以上)衍射光.此时在样品上形成的结构光不再是单一频率的余弦光栅图案,传统结构光显微成像中通过光栅3步相移成像获得层析图像的方法不再适用.新算法通过5步相移得到5幅源图像,也可重构出层析图像.结果表明,传统荧光图像中几乎不能分辨的某些精细结构,在重构的层析图像中能够清晰地分辨出来.%In our novel structured illumination microscope(SIM),a programmable digital micromirror device (DMD)was used in strad of piezo activeed grating to produce stable and reliable structured illumination. An optical low-pass spatial filter was applied to block the diffraction order higher than±Ist order. The illumination pattern was not a sinusoidal grid with a single spatial frequency, so the algorithm used in the traditional SIM is not proper here. AN improved algorithm based on the five raw images with different phase angles was introduced and discssed in detail. The results have demonstrated that the sectioned image can be reconstructed rom five raw images with different phase angles, with resolving some fine lateral structrue that was blurred in traditional microscopic image.