Super-resolution optical fluctuation imaging (SOFI) produces fast, background-free, super-resolved images by analyzing the temporal fluorescence fluctuations of independent emitters. With sufficient brightness and fluctuations, a higher order of image processing affords a higher resolution and in principle the resolution enhancement is unbounded. However, it is practically challenging to find suitable probes for high-order SOFI. Herein, we report two types of BODIPY-based polymer dots (Pdots) with narrow-band emissions, pronounced fluctuations, and prominent photostability, thus enabling high-order, dual-color SOFI nanoscopy. Single-particle and subcellular SOFI analysis reveals the superior performance of the BODIPY Pdots as compared to conventional streptavidin-conjugated Alexa dyes. In contrast with wide-field images, the spatial resolution (∼57 nm) was enhanced by ∼6.0-fold in 8^th-order single-particle SOFI nanoscopy. A spatial resolution (61 nm) was obtained for single microtubules labeled by the BODIPY Pdots, while the majority of the subcellular structures were lost for those labeled by streptavidin-Alexa dyes in 8^th-order SOFI. This work indicates the unprecedented performance of Pdot probes for multi-color subcellular SOFI applications.

中文翻译：

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具有明显荧光波动的窄带聚合物点，用于双色超分辨率成像

超分辨率光学波动成像（SOFI）通过分析独立发射器的时间荧光波动来生成快速，无背景的超分辨图像。具有足够的亮度和波动，较高级别的图像处理可提供更高的分辨率，并且原则上分辨率的提升是无限的。但是，为高阶SOFI寻找合适的探头实际上是一项挑战。在本文中，我们报告了两种类型的基于BODIPY的聚合物点（Pdot），它们具有窄带发射，明显的波动和突出的光稳定性，因此可以实现高阶双色SOFI纳米技术。单颗粒和亚细胞SOFI分析显示，与传统的链霉亲和素偶联的Alexa染料相比，BODIPY Pdots具有更好的性能。与广角图像相比，^个阶单粒子SOFI纳米显微镜。对于由P点BODIPY标记的单微管进行，得到空间分辨率（61纳米），而大多数的亚细胞结构的丢失对于那些由链霉亲和标记的Alexa染料在8^个阶SOFI。这项工作表明Pdot探针在多色亚细胞SOFI应用中具有空前的性能。