Choi Jong-ryul, Kim Donghyun
School of Electrical and Electronic Engineering, Yonsei University, Seoul, South Korea.
J Opt Soc Am A Opt Image Sci Vis. 2012 Oct 1;29(10):2165-73. doi: 10.1364/JOSAA.29.002165.
We investigate improved image reconstruction of structured light illumination for high-resolution imaging of three-dimensional (3D) cell-based assays. For proof of concept, an in situ fluorescence optical detection system was built with a digital micromirror device as a spatial light modulator, for which phase and tilting angle in a grid pattern were varied to implement specific image reconstruction schemes. Subtractive reconstruction algorithms based on structured light illumination were used to acquire images of fluorescent microbeads deposited as a two-dimensional monolayer or in 3D alginate matrix. We have confirmed that an optical subtraction algorithm improves axial and lateral resolution by effectively removing out-of-focus fluorescence. The results suggest that subtractive image reconstruction can be useful for structured illumination microscopy of broad types of cell-based assays with high image resolution.
我们研究了用于基于三维（3D）细胞分析的高分辨率成像的结构光照明的改进图像重建。为了验证概念，构建了一个原位荧光光学检测系统，该系统以数字微镜器件作为空间光调制器，其中以网格图案变化的相位和倾斜角来实现特定的图像重建方案。基于结构光照明的减法重建算法用于获取沉积为二维单层或三维藻酸盐基质中的荧光微珠的图像。我们已经证实，光学减法算法通过有效去除离焦荧光来提高轴向和横向分辨率。结果表明，减法图像重建对于具有高图像分辨率的广泛类型的基于细胞的分析的结构照明显微镜检查可能是有用的。
