Label-free difference super-resolution microscopy based on parallel detection

In this paper, a new method is proposed for super-resolution imaging of non-fluorescent samples. This approach is based on the intensity difference between confocal image and negative confocal image, which are simultaneously acquired at one sample scanning. In order to get these two different images simultaneously, the sample was illuminated by two different focused spots from the same laser source: the doughnut spot and the solid spot. The effectiveness of the label-free difference microscopy based on parallel detection was validated by experiments on some samples including 80 nm gold beads, 100 nm silver nanowires, and Blu-ray DVD without fluorescent dyes. By subtraction of the reflected light intensity from the sample, the final resolution of the image without deconvolution was enhanced about 1.6 times compared with confocal imaging. This technique can be applied to surface topography detection of metallographic or other non-fluorescent materials.

本文提出了一种用于非荧光样品超分辨率成像的新方法。该方法基于共聚焦图像和负共聚焦图像之间的强度差异，这两种图像是在一次样品扫描过程中同时获取的。为了同时获得这两种不同的图像，使用来自同一激光源的两个不同聚焦光斑照射样品：环形光斑和实心光斑。通过对包括80纳米金珠、100纳米银纳米线以及无荧光染料的蓝光DVD等一些样品进行实验，验证了基于并行检测的无标记差分显微技术的有效性。通过减去样品的反射光强度，未进行反卷积的图像最终分辨率与共聚焦成像相比提高了约1.6倍。该技术可应用于金相或其他非荧光材料的表面形貌检测。