Large Stokes-shift bioorthogonal probes for STED, 2P-STED and multi-color STED nanoscopy. (23rd December 2020)

• Record Type: Journal Article
• Authors: Török, György; Cserép, Gergely B; Telek, András; Arany, Dóra; Váradi, Melinda; Homolya, László; Kellermayer, Miklós; Kele, Péter; Németh, Krisztina
• Is Part Of: Materials research express. Volume 9:Number 1(2022)
• Publication Date: 2020-12-23
• Subjects: large Stokes-shift fluorescent dye -- bioorthogonal live-cell labeling -- genetic code expansion -- multi-color STED -- two-photon STED; Materials science -- Research -- Periodicals; Materials science -- Periodicals; 620.11
• DOI: 10.1088/2050-6120/abb363
• Languages: English
• ISSNs: 2053-1591

Abstract

Abstract: 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.