Molecular Tuning of Styryl Dyes Leads to Versatile and Efficient Plasma Membrane Probes for Cell and Tissue Imaging.,Bioconjugate Chemistry

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Molecular Tuning of Styryl Dyes Leads to Versatile and Efficient Plasma Membrane Probes for Cell and Tissue Imaging.
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2020-02-13 , DOI: 10.1021/acs.bioconjchem.0c00023
Mayeul Collot ₁ , Emmanuel Boutant ₁ , Kyong Tkhe Fam ₁ , Lydia Danglot ₂ , Andrey S Klymchenko ₁

Affiliation

The plasma membrane (PM) plays a major role in many biological processes; therefore, its proper fluorescence staining is required in bioimaging. Among the commercially available PM probes, styryl dye FM1-43 is one of the most widely used. In this work, we demonstrated that fine chemical modifications of FM1-43 can dramatically improve the PM staining. The newly developed probes, SP-468 and SQ-535, were found to display enhanced photophysical properties (reduced cross-talk, higher brightness, improved photostability) and, unlike FM1-43, provided excellent and immediate PM staining in 5 different mammalian cell types including neurons (primary culture and tissue imaging). Taking advantage of these features, we successfully used SP-468 in STED super resolution neuronal imaging. Additionally, we showed that the new probes displayed differences in their internalization pathways compared to their parent FM1-43. Finally, we showed that the new probes kept the ability to stain the PM of plant cells. Overall, this work presents new useful probes for PM imaging in cells and tissues and provides insights on the molecular design of new PM targeting molecules.

中文翻译：

苯乙烯基染料的分子调节可产生用于细胞和组织成像的多功能高效血浆膜探针。

质膜（PM）在许多生物学过程中都起着重要作用。因此，在生物成像中需要适当的荧光染色。在市售的PM探针中，苯乙烯基染料FM1-43是使用最广泛的探针之一。在这项工作中，我们证明了FM1-43的精细化学修饰可以显着改善PM染色。发现新开发的探针SP-468和SQ-535具有增强的光物理特性（减少的串扰，更高的亮度，改善的光稳定性），并且与FM1-43不同，它在5种不同的哺乳动物细胞中均具有出色的即时PM染色能力类型，包括神经元（原代培养和组织成像）。利用这些功能，我们成功地将SP-468用于STED超分辨率神经元成像。另外，我们发现，新探针与其母体FM1-43相比在其内部化途径上显示出差异。最后，我们证明了新探针具有对植物细胞PM染色的能力。总的来说，这项工作为细胞和组织中的PM成像提供了新的有用的探针，并为新型PM靶向分子的分子设计提供了见识。

更新日期：2020-02-27

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