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The development of new microbicidal compounds has become a top priority due to the emergence and spread of drug-resistant pathogenic microbes. In this study, blue-emitting and positively charged carbon dots (CDs), called Du-CDs, were fabricated for the first time utilizing the natural product extract of endophyte Diaporthe unshiuensis YSP3 as raw material through a one-step solvothermal method, which possessed varied functional groups including amino, carboxyl, hydroxyl, and sulfite groups. Interestingly, Du-CDs exhibited notably enhanced antimicrobial activities toward both bacteria and fungi as compared to the natural product extract of YSP3, with low minimum inhibitory concentrations. Moreover, Du-CDs significantly inhibited the formation of biofilms. Du-CDs bound with the microbial cell surface via electronic interaction or hydrophobic interaction entered the microbial cells and were distributed fully inside the cells. Du-CDs caused cell membrane damage and/or cell division cycle interruption, resulting in microbial cell death. Moreover, Du-CDs exhibited an improved antimicrobial effect and accelerated wound healing ability with good biocompatibility in the mouse model. Overall, we demonstrate that the formation of CDs from fungal natural products presents a promising and potential means to develop novel antimicrobial agents with great fluorescence, improved microbiocidal effect and wound healing capacity, and good biosafety for combating microbial infections.

Nucleus pH is closely linked to many diseases such as aging, heart disease, skeletal myopathies, cancer, Alzheimer's disease, etc. Nevertheless, fluorescent sensors that can directly monitor nucleus pH changes have not yet been reported. Here, we first reported a green emissive carbon dots (CDs) for nucleus pH detection in living cells. CDs can selectively target nucleus with high accumulation at nucleolus due to their high affinity towards RNA once entering cells by lipid raft mediated endocytosis. Without washing, CDs at 5 μg/mL was enough to lighten nucleus within 10 min with the fluorescence on ever after 24 h incubation, achieving fast, wash-free, and long-term nucleus/nucleolus imaging. Meanwhile, the luminescent intensity of CDs was reduced gradually when pH changed continuously from 1 to 12, showing a pH-responsive fluorescence property with two linear ranges of pH 2-7 and pH 7-12. With their nucleus-targeting ability and pH-dependent photoluminescent property, CDs was successfully leveraged for nucleus pH detection in A549 cells and for in vivo pH sensing in zebra fish. CDs present a promising and powerful fluorescent sensor for nucleus imaging and nucleus pH sensing in living cells on the way to understand nucleus-related biological events., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Given the significant impact of ions on environment pollution and human health, it is urgently needed to establish effective and convenient ion detection approaches, particularly in living cells. In this paper, we constructed multicolor N-doped-carbon dots (mPD-CDs) by facile one-step hydrothermal carbonization of m-phenylenediamine (mPD). mPD-CDs were successfully deployed for multicolor cellular imaging for animal cells, fungi, and bacteria in a wash-free way with high photostability and satisfactory biocompability. Moreover, mPD-CDs can be used as a fluorescent sensing probe for ultrasensitive detection of both iodide ion (I - ) and typical heavy metals such as cadmium (Cd 2+ ), copper (Cu 2+ ), mercury (Hg 2+ ), gadolinium (Gd 3+ ), ferrous ion (Fe 2+ ), Zinc (Zn 2+ ), and ferric ion (Fe 3+ ). This is the first report using CDs as optical sensing probe for the detection of Gd 3+ , and for detection of Fe 3+ with fluorescence "turn on". More significantly, with these versatile and fascinating properties, we applied mPD-CDs for intracellular ion detection in living cells like Hep G2 and S. cerevisiae, and zebra fish. Altogether, mPD-CDs displayed great potential for multicolor cell imaging and the multiple ion detection in vitro and in vivo, presenting a promising strategy for in-situ ultrasensitive sensing of multiple metal ions in the environment and the biological systems., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Congo red (CR) is a hazardous pigment, posing increasing dangerous to the environment and human health. However, the in-situ detection of CR in living cells has not been reported, as far as we know. Here, negatively-charged green-emitting Ca, N, S-doped carbon dots (Mis-mPD-CDs) were fabricated from plant and m-phenylenediamine (mPD) by facile one-step hydrothermal carbonization. Mis-mPD-CDs were capable of rapidly detecting CR on the basis of their fluorescence quenching by CR due to the inner filter effect. This CR detection based on Mis-mPD-CDs displayed a linear range of 0.2-1.2 μM and a low limit of detection (58 nM), and was not interfered by metal ions, important biological molecules, and other dyes, showing high sensitivity and selectivity. More interestingly, Mis-mPD-CDs can rapidly enter and label animal cells (A549, 4T1, and HUVEC), fungi (S. cerevisiae, C. albicans, and T. reesei), and bacteria (E. coli and S. aureus) for long term with high stability and appealing biocompatibility. Based on these compelling characteristics, we applied Mis-mPD-CDs for sensing and imaging CR in living cells (A549, C. albicans, E. coli, and S. aureus) and zebra fish. On the other hand, the quantitative detection of CR by Mis-mPD-CDs was realized in real samples like fish tissues and industrial wastewater. This is the first report on applying CDs for rapid CR detection in living cells and in vivo. Mis-mPD-CDs provides a novel efficient platform for probing intracellular CR, expanding the applications of CDs as biosensors for toxic dyes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The discrimination between dead and live cells is crucial for cell viability evaluation. Carbon dots (CDs), with advantages like simple and cost-effective synthesis, excellent biocompatibility, and high photostability, have shown potential for realizing selective live/dead cell staining. However, most of the developed CDs with the live/dead cell discrimination capacity usually have low photoluminescence quantum yields (PLQYs) and excitation wavelength-dependent fluorescence emission (which can cause fluorescence overlap with other fluorescent probes and make dual-color live/dead staining impossible), and hence, developing ultrabright CDs with excitation wavelength-independent fluorescence emission property for live/dead cell discrimination becomes an important task. Here, using a one-pot hydrothermal method, we prepared ultrasmall (∼1.6 nm), ultrabright (PLQY: ∼78%), and excitation wavelength-independent sulfur-doped carbon dots (termed S-CDs) using rose bengal and 1,4-dimercaptobenzene as raw materials and demonstrated that the S-CDs could rapidly (∼5 min) and accurately distinguish dead cells from live ones for almost all the cell types including bacterial, fungal, and animal cells in a wash-free manner. We confirmed that the S-CDs could rapidly pass through the dead cell surfaces to enter the interior of the dead cells, thus visualizing these dead cells. In contrast, the S-CDs could not enter the interior of live cells and thus could not stain these live cells. We further verified that the S-CDs presented better biocompatibility and higher photostability than the commercial live/dead staining dye propidium iodide, ensuring its bright application prospect in cell imaging and cell viability assessment. Overall, this work develops a type of CDs capable of realizing the live/dead cell discrimination of almost all the cell types (bacterial, fungal, and animal cells), which has seldom been achieved by other fluorescent nanoprobes.

Imaging biofilm-encased microorganisms is challenging due to them being shielded by a sticky and strong extracellular polymeric substance matrix that blocks the penetration of dyes, but is especially important for biofilm study. Here, carbon dots, namely CDs-605, were used for successfully imaging biofilm-encased microorganisms. We demonstrated that such CDs-605 could be easily synthesized from L. plantarum by one-step hydrothermal carbonization. The negatively charged CDs-605 were composed of C, H, O, N, P, and S and contained various functional groups, including -OH, -COOH, and -CONH-. For the first time, CDs-605 were found to be capable of imaging biofilm-encased microorganisms, including E. coli, S. oneidensis, P. aeruginosa, S. aures, and T. reesei, demonstrating that the labeling is universal. The labeling method based on CDs-605 did not require incubation, protection from light, or washing, which makes it very convenient. Additionally, CDs-605 did not disturb the biofilm and displayed better photostability compared to the commercial counterpart. CDs-605 can serve as a simple, yet powerful staining agent for biofilm-embedded microorganism imaging, which is vital for gaining a fundamental understanding of biofilm formation and dispersal.

Frequent bacterial/fungal infections and occurrence of antibiotic resistance pose increasing threats to the public and thus require the development of new antibacterial/antifungal agents and strategies. Carbon dots (CDs) have been well demonstrated to be promising and potent antimicrobial nanomaterials and serve as potential alternatives to conventional antibiotics. In recent years, great efforts have been made by many researchers to develop new carbon dot-based antimicrobial agents to combat microbial infections. Here, as an update to our previous relevant review (C 2019, 5, 33), we summarize the recent achievements in the utilization of CDs for microbial inactivation. We review four kinds of antimicrobial CDs including nitrogen-doped CDs, metal-containing CDs, antibiotic-conjugated CDs, and photoresponsive CDs in terms of their starting materials, synthetic route, surface functionalization, antimicrobial ability, and the related antimicrobial mechanism if available. In addition, we summarize the emerging applications of CD-related antimicrobial materials in medical and industry fields. Finally, we discuss the existing challenges of antimicrobial CDs and the future research directions that are worth exploring. We believe that this review provides a comprehensive overview of the recent advances in antimicrobial CDs and may inspire the development of new CDs with desirable antimicrobial activities. [ABSTRACT FROM AUTHOR]