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SCIENCE CHINA Materials, Volume 62 , Issue 10 : 1496-1504(2019) https://doi.org/10.1007/s40843-019-9450-3

Altering sub-cellular location for bioimaging by engineering the carbon based fluorescent nanoprobe

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  • ReceivedApr 15, 2019
  • AcceptedJun 3, 2019
  • PublishedJul 3, 2019

Abstract

碳基荧光纳米探针在生物成像领域展现出诱人的应用前景. 本文通过调节合成方法合成了一种磺酸基修饰的石墨烯量子点(S-GQDs)荧光探针. 该探针呈现出优异的光学和理化性能, 如荧光强度高、 pH稳定、表面带负电等. 研究表明其发光机理主要依赖荧光分子发光机制. 与我们之前报道的氨基化量子点A-GQDs和肿瘤细胞核靶向探针GTTN对比, 该探针具有良好的生物安全性, 可以在相当短的时间内即跨膜进入细胞, 而GTTN在正常的体外培养条件下无法进入细胞. 为此, 我们探究了产生该差异的原因. 结果表明, S-GQDs与A-GQDs截然不同的合成原料导致了他们的毒性差异, 而S-GQDs的不稳定性则是导致其进入细胞、与GTTN明显不同的主要原因.


Funded by

the National Natural Science Foundation of China(Nos.,21371115,11025526,1175107,21101104,11422542)

the Shanghai University-Universal Medical Imaging Diagnostic Research Foundation(19H00100)

the Program for Changjiang Scholars and Innovative Research Team in University(No.,IRT13078)


Acknowledgment

This work has been supported by the National Natural Science Foundation of China (21371115, 11025526, 1175107, 21101104 and 11422542), Shanghai University-Universal Medical Imaging Diagnostic Research Foundation (19H00100) and the Program for Changjiang Scholars and Innovative Research Team in University (IRT13078).


Interest statement

The authors declare that they have no conflict of interest.


Contributions statement

Zhang K, Yao C and Gao W designed and engineered the materials; Li C, Ding L and Huang Y conceived and performed the biological experiments and characterization of materials; Zhang J contributed to the discussion and paper writing. Wang Y convinced the idea. Wang Y and Wu M wrote the paper. All authors contributed to the general discussion.


Author information

Chenchen Li was born in 1991. She is a PhD student in Prof. Minghong Wu’s group and supervised by Prof. Yanli Wang. She joined Prof. Wu’s group as a PhD student in 2016. Her research interests focus on the biosecurity and bio-behavior of nanomaterials.


Yanli Wang obtained her PhD degree in environmental engineering from Shanghai University in 2010. Now she is the director of the Tumor Precision Targeting Research Center, Shanghai University, and a professor of the School of Environmental and Chemical Engineering, Shanghai University. Her main research interests include: 1. the application of intelligent targeted fluorescent nanomaterials in tumor diagnosis; 2. intelligent targeted nano-drug design and its application in tumor therapy; 3. the development of tumor marker detection kit; 4. biosecurity of nanomaterials.


Minghong Wu obtained her PhD degree from Shanghai Institute of Applied Physics of Chinese Academy of Sciences in 1999. She is the vice president of Shanghai University. She is the National Outstanding Youth, Yangtze River scholar of China and the foreign academicians of Russian Academy of Engineering and Russian Academy of Science. Her research interests mainly focus on bio-effects and safety evaluation of nanomaterials and environmental pollution analysis and control.


Supplement

Supplementary information

Experimental details and supporting data are available in the online version of the paper.


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  • Figure 1

    (a) Schematic illustration of the large-scale preparation steps of S-GQDs. (b) TEM image of S-GQDs (size distribution inset). Scale bar: 20 nm. (c) HRTEM image of S-GQDs. Scale bar: 1 nm; the lattice parameter is 0.24 nm. The inset is an FFT image of a corresponding area (selected by the yellow square). (d) The FT-IR spectra of S-GQDs. (e) The XRD pattern of S-GQDs. (f) XPS full survey of S-GQDs. The high resolution XPS of C 1s (g), O 1s (h), and S 2p (i).

  • Figure 2

    (a) UV-visible absorption (ABS), photoluminescent emission (PL) and photoluminescent excitation (PLE) spectra of S-GQDs. (b) PL spectra of S-GQDs excited at different excitation wavelengths varying from 360 to 460 nm. (c) Photostability of S-GQDs over a long period of time from one day to a month. (d) PL spectra of dialysate and remaining solution after S-GQDs dialysis. (e) Solvent-dependent PL intensity of S-GQDs collected at 373 nm excitation. (f) Solvent-dependent PL peaks of S-GQDs collected at 373 nm excitation.

  • Figure 3

    Confocal images of S-GQDs, A-GQDs and GTTN in 4T1 cell in vitro. (a) Cell membrane co-localization. (b) Lysosome co-localization. Cells were co-incubated with 100 mg L−1 S-GQDs, 30 mg L−1 A-GQDs or 100 mg L−1 GTTN for 48 h. Left, graphene-based nanoparticles (405 nm excitation); middle, cell membrane stained by CellMask Orange plasma membrane stain (561 nm excitation) or cell lysosome stained by LysoTracker Deep Red (635 nm excitation); right, merged image. Scale bar: 10 μm.

  • Figure 4

    Cell viability assays of (a) S-GQDs, (b) GTTN, and (c) A-GQDs at different concentrations for different incubation times detected by cck-8 assay. Representative light microscopic images of (d) S-GQDs of 300 mg L−1 (48 h, scale bar: 20 μm); (e) GTTN of 300 mg L−1 (48 h, scale bar: 20 μm); (f) A-GQDs of 50 mg L−1 (48 h, scale bar: 20 μm). Static settled S-GQDs and GTTN for different times from 5 min to 48 h in AS and culture medium. TEM images of S-GQDs (g) and GTTN (j) after being settled for 48 h. Scale bars: 200 nm (g); 100 nm (j). UV-visible absorption peak value changes of S-GQDs (h) and GTTN (k) at 398 nm. Photoluminescent peak value changes of S-GQDs (i) and GTTN (l) at 480 nm as the settling time increases.

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