Chowdhury Shwetadwip, Dhalla Al-Hafeez, Izatt Joseph
Department of Biomedical Engineering, Fitzpatrick Institute for Photonics, Duke University, 136 Hudson Hall, Durham NC 27708, USA.
Biomed Opt Express. 2012 Aug 1;3(8):1841-54. doi: 10.1364/BOE.3.001841. Epub 2012 Jul 12.
Many biological structures of interest are beyond the diffraction limit of conventional microscopes and their visualization requires application of super-resolution techniques. Such techniques have found remarkable success in surpassing the diffraction limit to achieve sub-diffraction limited resolution; however, they are predominantly limited to fluorescent samples. Here, we introduce a non-fluorescent analogue to structured illumination microscopy, termed structured oblique illumination microscopy (SOIM), where we use simultaneous oblique illuminations of the sample to multiplex high spatial-frequency content into the frequency support of the system. We introduce a theoretical framework describing how to demodulate this multiplexed information to reconstruct an image with a spatial-frequency support exceeding that of the system's classical diffraction limit. This approach allows enhanced-resolution imaging of non-fluorescent samples. Experimental confirmation of the approach is obtained in a reflection test target with moderate numerical aperture.
许多感兴趣的生物结构超出了传统显微镜的衍射极限，对其进行可视化需要应用超分辨率技术。这类技术在突破衍射极限以实现亚衍射极限分辨率方面取得了显著成功；然而，它们主要局限于荧光样本。在此，我们引入一种结构照明显微术的非荧光类似物，称为结构斜照明显微术（SOIM），我们利用对样本的同步斜照将高空间频率成分复用至系统的频率支持范围内。我们引入了一个理论框架，描述如何解调这种复用信息以重建空间频率支持超过系统经典衍射极限的图像。这种方法能够对非荧光样本进行增强分辨率成像。在具有中等数值孔径的反射测试靶标中获得了该方法的实验验证。
