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用于活细胞和组织中微管和气管的超分辨率显微镜观察的荧光染料和探针。

Fluorescent dyes and probes for super-resolution microscopy of microtubules and tracheoles in living cells and tissues.

作者信息

Lukinavičius Gražvydas, Mitronova Gyuzel Y, Schnorrenberg Sebastian, Butkevich Alexey N, Barthel Hannah, Belov Vladimir N, Hell Stefan W

机构信息

Max Planck Institute for Biophysical Chemistry , Department of NanoBiophotonics , Am Fassberg 11 , 37077 Göttingen , Germany . Email:

出版信息

Chem Sci. 2018 Feb 26;9(13):3324-3334. doi: 10.1039/c7sc05334g. eCollection 2018 Apr 7.

Abstract

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 () 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.

摘要

我们基于最近报道的荧光染料(510R、580CP、GeR和SiR)以及化疗药物——紫杉烷类(多西他赛、卡巴他赛和拉罗他赛),引入了荧光微管蛋白探针。最终探针的细胞毒性、染色性能和特异性强烈依赖于这两种成分。我们发现了聚集效率(与荧光团的螺内酯化有关)和细胞毒性之间的相关性。通过探针优化,我们在活的人类成纤维细胞微管网络的受激发射损耗(STED)显微镜图像中达到了29±11纳米的分辨率。应用于活的果蝇组织突出了两种不同的结构:微管和气管。我们将580CP和SiR染料的6-羧基异构体鉴定为含几丁质的气管螺旋丝(气管的一个组成部分)的标记物。STED显微镜揭示了气管螺旋丝的周期性与气管直径之间的相关性。微管蛋白和气管螺旋丝的联合STED成像显示了昆虫幼虫活组织中微管与呼吸网络之间的紧密相互作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb2/5932598/d66e973a6e49/c7sc05334g-f1.jpg

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