Fluorescent dyes and probes for super-resolution microscopy of microtubules and tracheoles in living cells and tissues.
Fluorescent dyes and probes for super-resolution microscopy of microtubules and tracheoles in living cells and tissues.
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DOI：
10.1039/c7sc05334g
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发表时间：
2018-04-07
期刊：
影响因子：
8.4
通讯作者：
Hell SW
中科院分区：
文献类型：
--
作者：
Lukinavičius G;Mitronova GY;Schnorrenberg S;Butkevich AN;Barthel H;Belov VN;Hell SW
纳米显微镜兼容荧光微管蛋白探针可用于染色昆虫气管内的微管和富含几丁质的带体。我们介绍了基于最近报道的荧光染料（510R, 580CP， GeR和SiR）和化疗药物紫杉烷（多西他赛，卡巴他赛和larotaxel）的荧光微管蛋白探针。最终探针的细胞毒性，其染色性能和特异性在很大程度上取决于这两种成分。我们发现聚集效率（与荧光团的旋内酯化有关）与细胞毒性之间存在相关性。探针优化使我们在受激发射损耗（STED）显微镜下获得的活体人成纤维细胞微管网络的分辨率达到29±11 nm。在活体果蝇（Drosophila melanogaster）组织中的应用突出了两种不同的结构：微管和气管。我们鉴定了580CP和SiR染料的6-羧基异构体作为气管成分含几丁质带绦虫的标记物。STED显微镜显示带绦虫的周期性与气管直径的相关性。微管蛋白和带绦虫STED联合成像显示，微管和呼吸网络在幼虫活体组织中有密切的相互作用。
Nanoscopy compatible fluorescent tubulin probes can be used to stain microtubules and chitin-rich taenidia in the insect tracheoles. We introduce fluorogenic tubulin probes based on the recently reported fluorescent dyes (510R, 580CP, GeR and SiR) and chemotherapy agents – taxanes (docetaxel, cabazitaxel and larotaxel). The cytotoxicity of the final probe, its staining performance and specificity strongly depend on both components. We found correlation between the aggregation efficiency (related to the spirolactonization of fluorophore) and cytotoxicity. Probe optimization allowed us to reach 29 ± 11 nm resolution in stimulated emission depletion (STED) microscopy images of the microtubule network in living human fibroblasts. Application to living fruit fly (Drosophila melanogaster) tissues highlighted two distinct structures: microtubules and tracheoles. We identified 6-carboxy isomers of 580CP and SiR dyes as markers for chitin-containing taenidia, a component of tracheoles. STED microscopy revealed correlation between the taenidia periodicity and the diameter of the tracheole. Combined tubulin and taenidia STED imaging showed close interaction between the microtubules and respiratory networks in living tissues of the insect larvae.
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影响因子：
120.1
作者：
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通讯作者：
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作者：
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通讯作者：
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