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一步合成两亲性碳点用于活细胞内质网的超分辨率成像

One-step synthesized amphiphilic carbon dots for the super-resolution imaging of endoplasmic reticulum in live cells.

作者信息

Li Jiajia, Zhang Longdi, Chen Juan, Zhang Ruilong, Liu Zhengjie, Zhao Jun, Liu Bianhua, Han Ming-Yong, Han Guangmei, Zhang Zhongping

机构信息

Information Materials and Intelligent Sensing Laboratory of Anhui Province, School of Chemistry and Chemical Engineering, Institutes of Physical Science and Information Technology, Anhui University Hefei 230601 P. R. China

Key Lab of Photovoltaic and Energy Conservation Materials, Institute of Solid State Physics, HFIPS, Chinese Academy of Sciences Hefei 230031 P. R. China.

出版信息

RSC Adv. 2022 Jul 5;12(30):19424-19430. doi: 10.1039/d2ra02705d. eCollection 2022 Jun 29.

Abstract

Stimulated emission depletion (STED) microscopy provides a powerful tool for visualizing the ultrastructure and dynamics of subcellular organelles, however, the photobleaching of organelle trackers have limited the application of STED imaging in living cells. Here, we report photostable and amphiphilic carbon dots (Phe-CDs) with bright orange fluorescence a simple one-pot hydrothermal treatment of -phenylenediamine and phenylalanine. The obtained Phe-CDs not only had high brightness (quantum yield ∼18%) but also showed excellent photostability under ultraviolet irradiation. The CDs can quickly penetrate into cells within 2 min and are specific for intracellular ER. The further investigations by Phe-CDs revealed the reconstitution process of ER from loosely spaced tubes into a continuously dense network of tubules and sheets during cell division. Importantly, compared with the standard microscopy, STED super-resolution imaging allowed the tracking of the ER ultrastructure with a lateral resolution less than 100 nm and the pores within the ER network are clearly visible. Moreover, the three dimensional (3D) structure of ER was also successfully reconstructed from z-stack images due to the excellent photostability of Phe-CDs.

摘要

受激辐射损耗(STED)显微镜为可视化亚细胞器的超微结构和动力学提供了一个强大的工具,然而,细胞器追踪剂的光漂白限制了STED成像在活细胞中的应用。在此,我们报道了通过对苯二胺和苯丙氨酸进行简单的一锅水热法处理制备的具有亮橙色荧光的光稳定两亲性碳点(Phe-CDs)。所制备的Phe-CDs不仅具有高亮度(量子产率约为18%),而且在紫外线照射下表现出优异的光稳定性。这些碳点能够在2分钟内快速穿透细胞,并且对细胞内的内质网具有特异性。利用Phe-CDs进行的进一步研究揭示了在细胞分裂过程中内质网从松散排列的管状结构重构为连续致密的管状和片状网络的过程。重要的是,与标准显微镜相比,STED超分辨率成像能够以小于100 nm的横向分辨率追踪内质网的超微结构,并且内质网网络中的孔隙清晰可见。此外,由于Phe-CDs具有优异的光稳定性,还成功地从z轴堆叠图像重建了内质网的三维(3D)结构。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615a/9255560/184bc6a66dd8/d2ra02705d-f1.jpg

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