Opt Express. 2021 Oct 11;29(21):34797-34809. doi: 10.1364/OE.440706.
Multi-color super-resolution localization microscopy (SRLM) provides great opportunities for studying the structural and functional details of biological samples. However, current multi-color SRLM methods either suffer from medium to high crosstalk, or require a dedicated optical system and a complicated image analysis procedure. To address these problems, here we propose a completely different method to realize multi-color SRLM. This method is built upon a customized RGBW camera with a repeated pattern of filtered (Red, Green, Blue and Near-infrared) and unfiltered (White) pixels. With a new insight that RGBW camera is advantageous for color recognition instead of color reproduction, we developed a joint encoding scheme of emitter location and color. By combing this RGBW camera with the joint encoding scheme and a simple optical set-up, we demonstrated two-color SRLM with ∼20 nm resolution and < 2% crosstalk (which is comparable to the best-reported values). This study significantly reduces the complexity of two-color SRLM (and potentially multi-color SRLM), and thus offers good opportunities for general biomedical research laboratories to use multi-color SRLM, which is currently mastered only by well-trained researchers.
多色超分辨率定位显微镜（SRLM）为研究生物样本的结构和功能细节提供了极好的机会。然而，现有的多色 SRLM 方法要么存在中等至高度的串扰，要么需要专用的光学系统和复杂的图像分析程序。为了解决这些问题，我们提出了一种完全不同的方法来实现多色 SRLM。该方法建立在具有滤色（红色、绿色、蓝色和近红外）和未滤色（白色）像素重复图案的定制 RGBW 相机上。基于 RGBW 相机在颜色识别方面优于颜色再现的新见解，我们开发了一种发射器位置和颜色的联合编码方案。通过将这种 RGBW 相机与联合编码方案和简单的光学设置相结合，我们展示了具有约 20nm 分辨率和<2%串扰（与最佳报道值相当）的双色 SRLM。这项研究显著降低了双色 SRLM（以及潜在的多色 SRLM）的复杂性，从而为一般生物医学研究实验室使用多色 SRLM 提供了良好的机会，而目前只有经过良好培训的研究人员才能掌握多色 SRLM。
