Synthesis and multiple STED imaging applications of four, red-emitting (610–670 nm), tetrazine-functionalized fluorescent probes (CBRD=Chemical Biology Research group Dye 1–4) with large Stokes-shift is presented. Present studies revealed the super-resolution microscopy applicability of the probes as demonstrated through bioorthogonal labeling scheme of cytoskeletal proteins actin and keratin-19, and mitochondrial protein TOMM20. Furthermore, super-resolved images of insulin receptors in live-cell bioorthogonal labeling schemes through a genetically encoded cyclooctynylated non-canonical amino acid are also presented. The large Stokes-shifts and the wide spectral bands of the probes enabled the use of two common depletion lasers (660 nm and 775 nm). The probes were also found suitable for super-resolution microscopy in combination with two-photon excitation (2P-STED) resulting in improved spatial resolution. One of the dyes was also used together with two commercial dyes in the three-color STED imaging of intracellular structures.

四种发红光（610-670 nm）四嗪功能化荧光探针的合成和多重 STED 成像应用（CBRD = 化学生物学研究组染料1-4) 具有大的斯托克斯位移。目前的研究揭示了探针的超分辨率显微镜适用性，如通过细胞骨架蛋白肌动蛋白和角蛋白 19 以及线粒体蛋白 TOMM20 的生物正交标记方案所证明的那样。此外，还展示了通过基因编码的环辛基化非规范氨基酸在活细胞生物正交标记方案中的胰岛素受体的超分辨率图像。探针的大斯托克斯位移和宽光谱带使得能够使用两种常见的耗尽激光器（660 nm 和 775 nm）。还发现这些探针适用于与双光子激发 (2P-STED) 相结合的超分辨率显微镜，从而提高了空间分辨率。