Differential labelling of human sub-cellular compartments with fluorescent dye esters and expansion microscopy.
Differential labelling of human sub-cellular compartments with fluorescent dye esters and expansion microscopy.
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DOI：
10.1039/d3nr01129a
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发表时间：
2023-11-23
期刊：
影响因子：
6.7
通讯作者：
Jayasinghe, Izzy
中科院分区：
文献类型：
--
作者：
Sheard, Thomas M. D.;Shakespeare, Tayla B.;Seehra, Rajpinder S.;Spencer, Michael E.;Suen, Kin M.;Jayasinghe, Izzy
芳香族荧光染料的胺反应性酯正在成为用于细胞和组织结构的非描述性染色的成像探针。我们的特点是染色模式的14个荧光染料酯物种不同的物理和光谱特性，在广泛研究的人HeLa细胞系。当与涉及可溶胀丙烯酰胺水凝胶的超分辨率技术膨胀显微镜（ExM）相结合时，荧光酯揭示了纳米级特征，包括细胞质膜结合的隔室和核仁密度。我们观察到的共轭染料的生化特性的差异标记模式。根据ExM方案中的应用时间点，观察到染色密度和隔室特异性的变化。检测到酯间相互作用引起的标记模式的额外复杂性。我们的研究结果提出了一些考虑使用荧光酯。我们证明了酯作为细胞蛋白质组染色剂库的有用补充，无论是单独应用以可视化整体细胞形态，还是作为与特定靶标记物如抗体一起提供超微结构背景的复染剂。使用超分辨率成像技术扩增显微镜显示14种不同疏水性和电荷的荧光NHS酯差异标记HeLa细胞中的一系列亚细胞区室。
Amine-reactive esters of aromatic fluorescent dyes are emerging as imaging probes for nondescript staining of cellular and tissue architectures. We characterised the staining patterns of 14 fluorescent dye ester species with varying physical and spectral properties in the broadly studied human HeLa cell line. When combined with the super-resolution technique expansion microscopy (ExM) involving swellable acrylamide hydrogels, fluorescent esters reveal nanoscale features including cytoplasmic membrane-bound compartments and nucleolar densities. We observe differential labelling patterns linked to the biochemical properties of the conjugated dye. Alterations in staining density and compartment specificity were seen depending on the timepoint of application in the ExM protocol. Additional complexity in labelling patterns was detected arising from inter-ester interactions. Our findings raise a number of considerations for the use of fluorescent esters. We demonstrate esters as a useful addition to the repertoire of stains of the cellular proteome, whether applied either on their own to visualise overall cellular morphology, or as counterstains providing ultrastructural context alongside specific target markers like antibodies. 14 fluorescent NHS esters of different hydrophobicity and charges were shown to differentially label a range of subcellular compartments in HeLa cells using the super-resolution imaging technique expansion microscopy.
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影响因子：
7.7
作者：
Yu, Chih-Chieh (Jay);Barry, Nicholas C.;Boyden, Edward S.
通讯作者：
Boyden, Edward S.
影响因子：
9.8
作者：
Bertiaux E;Balestra AC;Bournonville L;Louvel V;Maco B;Soldati-Favre D;Brochet M;Guichard P;Hamel V
通讯作者：
Hamel V
影响因子：
4.3
作者：
Kolmakov, Kirill;Belov, Vladimir N.;Hell, Stefan W.
通讯作者：
Hell, Stefan W.
影响因子：
3.2
作者：
Panchuk-Voloshina, N;Haugland, RP;Haugland, RP
通讯作者：
Haugland, RP
影响因子：
6.3
作者：
Rumin J;Bonnefond H;Saint-Jean B;Rouxel C;Sciandra A;Bernard O;Cadoret JP;Bougaran G
通讯作者：
Bougaran G