SIOC 中文
ACS
Adv search

Acta Chimica Sinica >

2020 , Vol. 78 >Issue 2: 130 - 139

DOI: https://doi.org/10.6023/A19100374

Article

A Reversibly Intramolecular Cyclization Cy5 Optical Probe for Stochastic Optical Reconstruction Microscopy in Live Cell Mitochondria

  • Yang Zhigang ,
  • Xiong Jia ,
  • Zhang Wei ,
  • Li Wen ,
  • Pan Wenhui ,
  • Zhang Jianguo ,
  • Gu Zhenyu ,
  • Huang Meina ,
  • Qu Junle
Expand
  • Center of Biomedical Photonics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060

Received date: 2019-10-19

  Online published: 2019-12-20

Supported by

Project supported by the National Natural Science Foundation of China (Nos. 61875131, 61525503) and Shenzhen Basic Research Project (Nos. JCYJ20170818100931714, JCYJ20180305125549234).

Fold

Abstract

Single molecule localization microscopy as an advanced optical imaging technique is capable of super-resolution imaging of biological targets with the size below the optical diffraction limit. It is promising to provide powerful tools for the exploration of occurrence mechanism of severe diseases and precisely therapeutic method at single cell/organelle level, which exhibits wide applications in biomedical field. Generally, stochastic optical reconstruction microscopy (STORM) is prominently dependent on large amount of imaging buffers (Redox enzymes) and thiol-containing reagents for the ideal photoblinking behaviors of optical probes. However, the imaging buffer and thiol-containing reagents are harmful for the live cells, which make it difficult to carry out STORM imaging in live cells. Therefore, it is of significance to exploit new approaches to display STORM imaging in live cells. In this work, we provided a new strategy to facilitate the design of live cell STORM imaging probes with improved photo-blinking mechanism. A new fluorescent pentamethine cyanine probe with a thiol-attachment (SHCH2CH2CH2-) at the N-position of one indoline moiety was synthesized to show spontaneously photoblinking behavior caused by intramolecular ring-closing/-opening processes. The fluorescent probe is biologically compatible with rare cytotoxicity and suitable for the live cell imaging. The probe can exhibit excellent photo-blinking under the direct illumination of a single laser beam (656 nm) with low power density (200 W·cm-2 for solution sample and 100 W·cm-2 for cell sample, respectively), without using any imaging buffer or thiol-chemicals. And the fluorescent probe was used to test cell toxicity with CCK-8, showed almost no cytotoxicity after 24 h incubation. The photo-blinking frames were collected with an electron multiplying charge coupled device (EMCCD, 60 Hz), and different frames were used to pre-treat with ImageJ software and then reconstruct STORM images with a Falcon algorithm to show marked imaging resolution enhancement, compared with wide-field images, which provide a new protocol for biomedical imaging.

Key words： single-molecule localization microscopy; super-resolution imaging; pentamethine cyanine; photo-blinking; mitochondrial probe

Cite this article

Yang Zhigang , Xiong Jia , Zhang Wei , Li Wen , Pan Wenhui , Zhang Jianguo , Gu Zhenyu , Huang Meina , Qu Junle . A Reversibly Intramolecular Cyclization Cy5 Optical Probe for Stochastic Optical Reconstruction Microscopy in Live Cell Mitochondria[J]. Acta Chimica Sinica, 2020 , 78(2) : 130 -139 . DOI: 10.6023/A19100374

References

[1] Yang, Z.; Cao, J.; He, Y.; Yang, J. H.; Kim, T.; Peng, X.; Kim, J. S. Chem. Soc. Rev. 2014, 43, 4563.
[2] Yang, Z.; Lee, J. H.; Jeon, H. M.; Han, J. H.; Park, N.; He, Y.; Lee, H.; Hong, K. S.; Kang, C.; Kim, J. S. J. Am. Chem. Soc. 2013, 135, 11657.
[3] Yang, Z.; He, Y.; Lee, J.-H.; Park, N.; Suh, M.; Chae, W.-S.; Cao, J.; Peng, X.; Jung, H.; Kang, C.; Kim, J. S. J. Am. Chem. Soc. 2013, 135, 9181.
[4] Lee, M. H.; Yang, Z.; Lim, C. W.; Lee, Y. H.; Dongbang, S.; Kang, C.; Kim, J. S. Chem. Rev. 2013, 113, 5071.
[5] Peng, X.; Yang, Z.; Wang, J.; Fan, J.; He, Y.; Song, F.; Wang, B.; Sun, S.; Qu, J.; Qi, J.; Yan, M. J. Am. Chem. Soc. 2011, 133, 6626.
[6] Huang, C.; Chen, H.; Li, F. Chin. J. Org. Chem. 2019, 39, 2467(in Chinese). (黄池宝, 陈会, 李福琴, 安思雅, 有机化学, 2019, 39, 2467.)
[7] Ju, Z.; Shu, P.; Xie, Z. Chin. J. Org. Chem. 2019, 39, 697(in Chinese). (鞠志宇, 舒朋华, 谢智宇, 蒋雨晴, 陶伟杰, 许志红, 有机化学, 2019, 39, 697.)
[8] Xie, Z.; Fu, M.; Yin, B. Chin. J. Org. Chem. 2018, 38, 1364(in Chinese). (谢振达, 付曼琳, 尹彪, 朱勍, 有机化学, 2018, 38, 1364.)
[9] Xi, Z.; Yuan, F.; Wang, Z. Acta Chim. Sinica 2018, 76, 460(in Chinese). (郗梓帆, 袁方龙, 王子飞, 李淑花, 范楼珍, 化学学报, 2018, 76, 460.)
[10] Xu, Y.; Zhao, Y.; Zhang, Y. Acta Chim. Sinica 2018, 76, 393(in Chinese). (徐毅, 赵彦, 张叶俊, 崔之芬, 王丽华, 樊春海, 高基民, 孙艳红, 化学学报, 2018, 76, 393.)
[11] Gu, T.; Shi, J.; Hua, Y. Acta Chim. Sinica 2017, 75, 991(in Chinese). (顾天航, 石君明, 滑熠龙, 刘静, 王伟, 张伟贤, 化学学报, 2017, 75, 991.)
[12] Wang, Y.; Wang, W. Acta Chim. Sinica 2017, 75, 1061(in Chinese). (王咏婕, 王伟, 化学学报, 2017, 75, 1061.)
[13] Yang, L.; Liu, B.; Li, N. Acta Chim. Sinica 2017, 75, 1047(in Chinese). (杨立敏, 刘波, 李娜, 唐波, 化学学报, 2017, 75, 1047.)
[14] Abbe, E. Arch. Mikroskop Anat. 1873, 9, 413.
[15] Yang, Z.; Sharma, A.; Qi, J.; Peng, X.; Lee, D. Y.; Hu, R.; Lin, D.; Qu, J.; Kim, J. S. Chem. Soc. Rev. 2016, 45, 4651.
[16] Huang, B.; Wang, W.; Bates, M.; Zhuang, X. Science 2008, 319, 807.
[17] Samanta, S.; He, Y.; Sharma, A.; Kim, J.; Pan, W.; Yang, Z.; Li, J.; Yan, W.; Liu, L.; Qu, J.; Kim, J. S. CHEM 2019, 5, 1697.
[18] Dempsey, G. T.; Bates, M.; Kowtoniuk, W. E.; Liu, D. R.; Tsien, R. Y.; Zhuang, X. J. Am. Chem. Soc. 2009, 131, 18192.
[19] Dempsey, G. T.; Vaughan, J. C.; Chen, K. H.; Bates, M.; Zhuang, X. Nat. Methods 2011, 8, 1027.
[20] Uno, S.; Kamiya, M.; Yoshihara, T.; Sugawara, K.; Okabe, K.; Tarhan, M. C.; Fujita, H.; Funatsu, T.; Okada, Y.; Tobita S.; Urano, Y. Nat. Chem. 2014, 6, 681.
[21] Samanta, S.; Gong, W.; Li, W.; Sharma, A.; Shim, I.; Zhang, W.; Das, P.; Pan, W.; Liu, L.; Yang, Z.; Qu, J.; Kim, J. S. Coord. Chem. Rev. 2019, 380, 17.
[22] Hell, S. W. Science 2007, 316, 1153.
[23] Huang, B.; Bates, M.; Zhuang, X. Ann. Rev. Biochem. 2009, 78, 993.
[24] Betzig, E.; Patterson, G. H.; Sougrat, R.; Lindwasser, O. W.; Olenych, S.; Bonifacino, J. S.; Davidson, M. W.; Lippincott-Schwartz, J.; Hess, H. F. Science 2006, 313, 5793.
[25] Rust, M. J.; Bates, M.; Zhuang, X. Nat. Methods 2006, 3, 793.
[26] Dertinger, T.; Colyer, R.; Iyer, G.; Weiss, S.; Enderlein, J. Proc. Natl. Acad. Sci. 2009, 106, 22287.
[27] Bates, M.; Huang, B.; Dempsey, G. T.; Zhuang, X. Science 2007, 317, 1749.
[28] Mortensen, K. I.; Churchman, L. S.; Spudich, J. A.; Flyvbjerg, H. Nat. Methods 2010, 7, 3778.
[29] Huang, B.; Babcock, H.; Zhuang, X. Cell 2010, 143, 1047.
[30] Miki, K.; Kojima, K.; Oride, K.; Harada, H.; Morinibu, A.; Ohe, K. Chem. Commun. 2017, 53, 7792.
[31] Min, J.; Vonesch, C.; Kirshner, H.; Carlini, L.; Olivier, N.; Holden, S.; Manley, S.; Ye, J. C.; Unser, M. Sci. Rep. 2014, 4, 4577.
Options
Outlines

/

〈 〉