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

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


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)


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.


Supplementary information

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


[1] Schroeder KL, Goreham RV, Nann T. Graphene quantum dots for theranostics and bioimaging. Pharm Res, 2016, 33: 2337-2357 CrossRef PubMed Google Scholar

[2] Kumar V, Singh V, Umrao S, et al. Facile, rapid and upscaled synthesis of green luminescent functional graphene quantum dots for bioimaging. RSC Adv, 2014, 4: 21101 CrossRef Google Scholar

[3] Du Y, Guo S. Chemically doped fluorescent carbon and graphene quantum dots for bioimaging, sensor, catalytic and photoelectronic applications. Nanoscale, 2016, 8: 2532-2543 CrossRef PubMed ADS Google Scholar

[4] Zhu S, Zhang J, Qiao C, et al. Strongly green-photoluminescent graphene quantum dots for bioimaging applications. Chem Commun, 2011, 47: 6858-6860 CrossRef PubMed Google Scholar

[5] Wang L, Li W, Wu B, et al. Facile synthesis of fluorescent graphene quantum dots from coffee grounds for bioimaging and sensing. Chem Eng J, 2016, 300: 75-82 CrossRef Google Scholar

[6] Zeng X, Bao J, Han M, et al. Quantum dots sensitized titanium dioxide decorated reduced graphene oxide for visible light excited photoelectrochemical biosensing at a low potential. Biosens, 2014, 54: 331-338 CrossRef PubMed Google Scholar

[7] Chen J, Zhao M, Li Y, et al. Synthesis of reduced graphene oxide intercalated ZnO quantum dots nanoballs for selective biosensing detection. Appl Surf Sci, 2016, 376: 133-137 CrossRef ADS Google Scholar

[8] Liu Z, Robinson JT, Tabakman SM, et al. Carbon materials for drug delivery & cancer therapy. Mater Today, 2011, 14: 316-323 CrossRef Google Scholar

[9] Tang J, Kong B, Wu H, et al. Carbon nanodots featuring efficient FRET for real-time monitoring of drug delivery and two-photon imaging. Adv Mater, 2013, 25: 6569-6574 CrossRef PubMed Google Scholar

[10] Hola K, Zhang Y, Wang Y, et al. Carbon dots—Emerging light emitters for bioimaging, cancer therapy and optoelectronics. Nano Today, 2014, 9: 590-603 CrossRef Google Scholar

[11] Peng Z, Han X, Li S, et al. Carbon dots: Biomacromolecule interaction, bioimaging and nanomedicine. Coord Chem Rev, 2017, 343: 256-277 CrossRef Google Scholar

[12] Wang H, Chen Q, Zhou S. Carbon-based hybrid nanogels: A synergistic nanoplatform for combined biosensing, bioimaging, and responsive drug delivery. Chem Soc Rev, 2018, 47: 4198-4232 CrossRef PubMed Google Scholar

[13] Zhang Q, Jie J, Diao S, et al. Solution-processed graphene quantum dot deep-UV photodetectors. ACS Nano, 2015, 9: 1561-1570 CrossRef Google Scholar

[14] Dong Y, Dai R, Dong T, et al. Photoluminescence, chemiluminescence and anodic electrochemiluminescence of hydrazide-modified graphene quantum dots. Nanoscale, 2014, 6: 11240-11245 CrossRef PubMed ADS Google Scholar

[15] Chiang CW, Haider G, Tan WC, et al. Highly stretchable and sensitive photodetectors based on hybrid graphene and graphene quantum dots. ACS Appl Mater Interfaces, 2016, 8: 466-471 CrossRef Google Scholar

[16] Yan J, Ye Q, Wang X, et al. CdS/CdSe quantum dot co-sensitized graphene nanocomposites via polymer brush templated synthesis for potential photovoltaic applications. Nanoscale, 2012, 4: 2109-2116 CrossRef PubMed ADS Google Scholar

[17] Ma WL, Li SS. Electrically controllable energy gaps in graphene quantum dots. Appl Phys Lett, 2012, 100: 163109 CrossRef ADS Google Scholar

[18] Fan Z, Li S, Yuan F, et al. Fluorescent graphene quantum dots for biosensing and bioimaging. RSC Adv, 2015, 5: 19773-19789 CrossRef Google Scholar

[19] Markovic ZM, Ristic BZ, Arsikin KM, et al. Graphene quantum dots as autophagy-inducing photodynamic agents. Biomaterials, 2012, 33: 7084-7092 CrossRef PubMed Google Scholar

[20] Amjadi M, Manzoori JL, Hallaj T. Chemiluminescence of graphene quantum dots and its application to the determination of uric acid. J Lumin, 2014, 153: 73-78 CrossRef ADS Google Scholar

[21] Kanodarwala FK, Wang F, Reece PJ, et al. Deposition of CdSe quantum dots on graphene sheets. J Lumin, 2014, 146: 46-52 CrossRef ADS Google Scholar

[22] Antonova IV, Nebogatikova NA, Prinz VY. Fluorinated graphene films with graphene quantum dots for electronic applications. J Appl Phys, 2016, 119: 224302 CrossRef ADS Google Scholar

[23] Lu L, Zhu Y, Shi C, et al. Large-scale synthesis of defect-selective graphene quantum dots by ultrasonic-assisted liquid-phase exfoliation. Carbon, 2016, 109: 373-383 CrossRef Google Scholar

[24] Chen Q, Shi C, Zhang C, et al. Magnetic enhancement of photoluminescence from blue-luminescent graphene quantum dots. Appl Phys Lett, 2016, 108: 061904 CrossRef ADS Google Scholar

[25] Jiang SD, Tang G, Ma YF, et al. Synthesis of nitrogen-doped graphene–ZnS quantum dots composites with highly efficient visible light photodegradation. Mater Chem Phys, 2015, 151: 34-42 CrossRef Google Scholar

[26] Zhang RX, Cai P, Zhang T, et al. Polymer–lipid hybrid nanoparticles synchronize pharmacokinetics of co-encapsulated doxorubicin–mitomycin C and enable their spatiotemporal co-delivery and local bioavailability in breast tumor. NanoMed-Nanotechnol Biol Med, 2016, 12: 1279-1290 CrossRef PubMed Google Scholar

[27] Sun L, Zhou X, Zhang Y, et al. Enhanced field emission of graphene–ZnO quantum dots hybrid structure. J Alloys Compd, 2015, 632: 604-608 CrossRef Google Scholar

[28] Pham CV, Madsuha AF, Nguyen TV, et al. Graphene-quantum dot hybrid materials on the road to optoelectronic applications. Synth Met, 2016, 219: 33-43 CrossRef Google Scholar

[29] Li Y, Hu Y, Zhao Y, et al. An electrochemical avenue to green-luminescent graphene quantum dots as potential electron-acceptors for photovoltaics. Adv Mater, 2011, 23: 776-780 CrossRef PubMed Google Scholar

[30] Mombrú D, Romero M, Faccio R, et al. Tuning electrical transport mechanism of polyaniline–graphene oxide quantum dots nanocomposites for potential electronic device applications. J Phys Chem C, 2016, 120: 25117-25123 CrossRef Google Scholar

[31] Liang L, Kong Z, Kang Z, et al. Theoretical evaluation on potential cytotoxicity of graphene quantum dots. ACS Biomater Sci Eng, 2016, 2: 1983-1991 CrossRef Google Scholar

[32] Jin Z, Owour P, Lei S, et al. Graphene, graphene quantum dots and their applications in optoelectronics. Curr Opin Colloid Interface Sci, 2015, 20: 439-453 CrossRef Google Scholar

[33] Lim CS, Hola K, Ambrosi A, et al. Graphene and carbon quantum dots electrochemistry. Electrochem Commun, 2015, 52: 75-79 CrossRef Google Scholar

[34] Dong Y, Tian W, Ren S, et al. Graphene quantum dots/L-cysteine coreactant electrochemiluminescence system and its application in sensing lead(II) ions. ACS Appl Mater Interfaces, 2014, 6: 1646-1651 CrossRef PubMed Google Scholar

[35] Justin R, Tao K, Román S, et al. Photoluminescent and superparamagnetic reduced graphene oxide–iron oxide quantum dots for dual-modality imaging, drug delivery and photothermal therapy. Carbon, 2016, 97: 54-70 CrossRef Google Scholar

[36] Wang L, Zhu SJ, Wang HY, et al. Common origin of green luminescence in carbon nanodots and graphene quantum dots. ACS Nano, 2014, 8: 2541-2547 CrossRef PubMed Google Scholar

[37] Roy P, Periasamy AP, Chuang C, et al. Plant leaf-derived graphene quantum dots and applications for white LEDs. New J Chem, 2014, 38: 4946-4951 CrossRef Google Scholar

[38] Yan L, Zhang Y, Zhang X, et al. Single layer graphene electrodes for quantum dot-light emitting diodes. Nanotechnology, 2015, 26: 135201 CrossRef PubMed ADS Google Scholar

[39] Zhu G, Xu T, Lv T, et al. Graphene-incorporated nanocrystalline TiO2 films for CdS quantum dot-sensitized solar cells. J Electroanal Chem, 2011, 650: 248-251 CrossRef Google Scholar

[40] Zhu Z, Ma J, Wang Z, et al. Efficiency enhancement of perovskite solar cells through fast electron extraction: The role of graphene quantum dots. J Am Chem Soc, 2014, 136: 3760-3763 CrossRef PubMed Google Scholar

[41] Zhuo S, Shao M, Lee ST. Upconversion and downconversion fluorescent graphene quantum dots: Ultrasonic preparation and photocatalysis. ACS Nano, 2012, 6: 1059-1064 CrossRef PubMed Google Scholar

[42] Li Y, Chopra N. Fabrication of nanoscale heterostructures comprised of graphene-encapsulated gold nanoparticles and semiconducting quantum dots for photocatalysis. Phys Chem Chem Phys, 2015, 17: 12881-12893 CrossRef PubMed ADS Google Scholar

[43] Tetsuka H, Nagoya A, Asahi R. Highly luminescent flexible amino-functionalized graphene quantum dots@cellulose nanofiber–clay hybrids for white-light emitting diodes. J Mater Chem C, 2015, 3: 3536-3541 CrossRef Google Scholar

[44] Wang L, Wang Y, Xu T, et al. Gram-scale synthesis of single-crystalline graphene quantum dots with superior optical properties. Nat Commun, 2014, 5: 5357 CrossRef PubMed ADS Google Scholar

[45] Zhu S, Song Y, Zhao X, et al. The photoluminescence mechanism in carbon dots (graphene quantum dots, carbon nanodots, and polymer dots): Current state and future perspective. Nano Res, 2015, 8: 355-381 CrossRef Google Scholar

[46] Zhou L, Geng J, Liu B. Graphene quantum dots from polycyclic aromatic hydrocarbon for bioimaging and sensing of Fe3+ and hydrogen peroxide. Part Part Syst Charact, 2013, 30: 1086-1092 CrossRef Google Scholar

[47] Wu X, Tian F, Wang W, et al. Fabrication of highly fluorescent graphene quantum dots using L-glutamic acid for in vitro/in vivo imaging and sensing. J Mater Chem C, 2013, 1: 4676-4684 CrossRef PubMed Google Scholar

[48] Song Y, Zhu S, Zhang S, et al. Investigation from chemical structure to photoluminescent mechanism: A type of carbon dots from the pyrolysis of citric acid and an amine. J Mater Chem C, 2015, 3: 5976-5984 CrossRef Google Scholar

[49] Shi F, Zhang Y, Na W, et al. Graphene quantum dots as selective fluorescence sensor for the detection of ascorbic acid and acid phosphatase via Cr(VI)/Cr(III)-modulated redox reaction. J Mater Chem B, 2016, 4: 3278-3285 CrossRef Google Scholar

[50] Lei Z, Ding L, Yao C, et al. A highly efficient tumor-targeting nanoprobe with a novel cell membrane permeability mechanism. Adv Mater, 2019, 31: 1807456 CrossRef PubMed Google Scholar

[51] Pan D, Guo L, Zhang J, et al. Cutting sp2 clusters in graphene sheets into colloidal graphene quantum dots with strong green fluorescence. J Mater Chem, 2012, 22: 3314-3318 CrossRef Google Scholar

[52] Pan D, Zhang J, Li Z, et al. Hydrothermal route for cutting graphene sheets into blue-luminescent graphene quantum dots. Adv Mater, 2010, 22: 734-738 CrossRef PubMed Google Scholar

[53] Xu H, Zhou S, Xiao L, et al. Fabrication of a nitrogen-doped graphene quantum dot from MOF-derived porous carbon and its application for highly selective fluorescence detection of Fe3+. J Mater Chem C, 2015, 3: 291-297 CrossRef Google Scholar

[54] Zhu S, Zhang J, Tang S, et al. Surface chemistry routes to modulate the photoluminescence of graphene quantum dots: From fluorescence mechanism to up-conversion bioimaging applications. Adv Funct Mater, 2012, 22: 4732-4740 CrossRef Google Scholar

[55] Kim S, Hwang SW, Kim MK, et al. Anomalous behaviors of visible luminescence from graphene quantum dots: Interplay between size and shape. ACS Nano, 2012, 6: 8203-8208 CrossRef PubMed Google Scholar

[56] He P, Sun J, Tian S, et al. Processable aqueous dispersions of graphene stabilized by graphene quantum dots. Chem Mater, 2014, 27: 218-226 CrossRef Google Scholar

[57] Canton I, Battaglia G. Endocytosis at the nanoscale. Chem Soc Rev, 2012, 41: 2718-2739 CrossRef PubMed Google Scholar

[58] Liu H, Sun Y, Li Z, et al. Lysosome-targeted carbon dots for ratiometric imaging of formaldehyde in living cells. Nanoscale, 2019, 11: 8458-8463 CrossRef PubMed Google Scholar

[59] Stern ST, Adiseshaiah PP, Crist RM. Autophagy and lysosomal dysfunction as emerging mechanisms of nanomaterial toxicity. Part Fibre Toxicol, 2012, 9: 20 CrossRef PubMed Google Scholar

[60] Lalwani G, D'Agati M, Khan AM, et al. Toxicology of graphene-based nanomaterials. Adv Drug Deliver Rev, 2016, 105: 109-144 CrossRef PubMed Google Scholar

[61] Singh H, Sreedharan S, Tiwari K, et al. Two photon excitable graphene quantum dots for structured illumination microscopy and imaging applications: Lysosome specificity and tissue-dependent imaging. Chem Commun, 2019, 55: 521-524 CrossRef PubMed Google Scholar

[62] Khlebtsov N, Dykman L. Biodistribution and toxicity of engineered gold nanoparticles: A review of in vitro and in vivo studies. Chem Soc Rev, 2011, 40: 1647-1671 CrossRef PubMed Google Scholar

[63] Rejman J, Nazarenus M, Jimenez de Aberasturi D, et al. Some thoughts about the intracellular location of nanoparticles and the resulting consequences. J Colloid Interface Sci, 2016, 482: 260-266 CrossRef PubMed ADS Google Scholar

[64] He B, Shi Y, Liang Y, et al. Single-walled carbon-nanohorns improve biocompatibility over nanotubes by triggering less protein-initiated pyroptosis and apoptosis in macrophages. Nat Commun, 2018, 9: 2393 CrossRef PubMed ADS Google Scholar

[65] Zhang QQ, Yang T, Li RS, et al. A functional preservation strategy for the production of highly photoluminescent emerald carbon dots for lysosome targeting and lysosomal pH imaging. Nanoscale, 2018, 10: 14705-14711 CrossRef PubMed Google Scholar

[66] Fiandra L, Mazzucchelli S, De Palma C, et al. Assessing the in vivo targeting efficiency of multifunctional nanoconstructs bearing antibody-derived ligands. ACS Nano, 2013, 7: 6092-6102 CrossRef PubMed Google Scholar

[67] Palevsky PM. Unresolved issues in dialysis: dialysis modality and dosing strategy in acute renal failure. Seminars Dialysis, 2006, 19: 165-170 CrossRef PubMed Google Scholar

  • 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.

Copyright 2020  CHINA SCIENCE PUBLISHING & MEDIA LTD.  中国科技出版传媒股份有限公司  版权所有

京ICP备14028887号-23       京公网安备11010102003388号