Theranostics 2020; 10(13):6072-6081. doi:10.7150/thno.42134 This issue Cite

Research Paper

Super-resolution observation of lysosomal dynamics with fluorescent gold nanoparticles

Kangqiang Qiu1,2, Yang Du2,3, Jiyan Liu3, Jun-Lin Guan2, Hui Chao1✉, Jiajie Diao2✉

1. MOE Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry, Sun Yat-Sen University, Guangzhou 510275, China.
2. Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA.
3. Department of Biotherapy, Cancer Center, State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China.

Citation:
Qiu K, Du Y, Liu J, Guan JL, Chao H, Diao J. Super-resolution observation of lysosomal dynamics with fluorescent gold nanoparticles. Theranostics 2020; 10(13):6072-6081. doi:10.7150/thno.42134. https://www.thno.org/v10p6072.htm
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Abstract

Graphic abstract

Because lysosomes play critical roles in multiple cellular functions and are associated with many diseases, studying them at the subcellular level could elucidate their functionality and support the discovery of therapeutic drugs for treating those diseases. The commonly used dyes for super-resolution imaging of lysosomes are the commercial molecular LysoTrackers. But the tolerance to changes in the lysosomal microenvironment and to lysosomal membrane permeabilization (LMP) and the photostability of the LysoTrackers are worrisome. The purpose of our study was to evaluate the feasibility of performing a fluorescent gold nanoprobe for super-resolution observation of lysosomal dynamics in living cells and compare it to the commercial LysoTrackers.

Methods: The nanoprobe Cy5@Au NP contained three parts: a bio-inert gold core, a biocompatible polyethylene glycol spacer, and a fluorophore cyanine 5. Structured illumination microscopy (SIM) was employed to capture the fluorescence of Cy5@Au NPs in cells. The tolerance assays to changes in the lysosomal microenvironment and to LMP, the photobleaching assay, and the long-term lysosomes labelling assay of Cy5@Au NPs were compared with commercial LysoTrackers. The super-resolution observation of lysosomal dynamics with Cy5@Au NPs was performed.

Results: Cy5@Au NPs can light up lysosomes specifically under SIM. Compared with commercial lysosomal molecular probes, Cy5@Au NPs exhibited stronger tolerance in lysosomes during various treatments, and changes in the lysosomal microenvironment and LMP did not cause Cy5@Au NPs to lose track of their targets. Cy5@Au NPs demonstrated an excellent anti-photobleaching ability, and a long-term labelling assay revealed that they could label lysosomes more than 3 d. Biological events of lysosomes such as the kiss-and-run process, fusion, fission, and mitophagy were recorded with the fluorescent Cy5@Au NPs under SIM.

Conclusions: The nanoprobe Cy5@Au NP was successfully used as a lysosomal probe for the super-resolution observation in living cells and found to overcome the limitations of commercial LysoTrackers. Our results thus confirm that nanoparticles can be useful tools for subcellular super-resolution imaging and highlight new avenues for using nanoparticles in biology.

Keywords: super-resolution imaging, structured illumination microscopy, lysosomes, mitophagy, long-term labelling


Citation styles

APA
Qiu, K., Du, Y., Liu, J., Guan, J.L., Chao, H., Diao, J. (2020). Super-resolution observation of lysosomal dynamics with fluorescent gold nanoparticles. Theranostics, 10(13), 6072-6081. https://doi.org/10.7150/thno.42134.

ACS
Qiu, K.; Du, Y.; Liu, J.; Guan, J.L.; Chao, H.; Diao, J. Super-resolution observation of lysosomal dynamics with fluorescent gold nanoparticles. Theranostics 2020, 10 (13), 6072-6081. DOI: 10.7150/thno.42134.

NLM
Qiu K, Du Y, Liu J, Guan JL, Chao H, Diao J. Super-resolution observation of lysosomal dynamics with fluorescent gold nanoparticles. Theranostics 2020; 10(13):6072-6081. doi:10.7150/thno.42134. https://www.thno.org/v10p6072.htm

CSE
Qiu K, Du Y, Liu J, Guan JL, Chao H, Diao J. 2020. Super-resolution observation of lysosomal dynamics with fluorescent gold nanoparticles. Theranostics. 10(13):6072-6081.

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