We report that nanoscale carbon particles (carbon dots) upon simple surface passivation are strongly photoluminescent in both solution and the solid state. The luminescence emission of the carbon dots is stable against photobleaching, and there is no blinking effect. These strongly emissive carbon dots may find applications similar to or beyond those of their widely pursued silicon counterparts.
Two-photon fluorescence materials have attracted much recent attention for their many promising applications, especially in the growing field of biomedical imaging. 1-5 Among the best performing two-photon fluorescence materials are semiconductor quantum dots such as CdSe and related core-shell nanoparticles. 6-8 These quantum dots have been demonstrated in various optical imaging experiments in vitro and in vivo. 6,9,10 At the same time, however, heavy metals as the essential elements in available high-performance semiconductor quantum dots have prompted serious health and environmental concerns in the community and beyond. Therefore, the search for benign alternatives has become increasingly important and urgent. Recently, we found and reported 11 that nanosized pure carbon particles may be surfacepassivated to exhibit bright photoluminescence in the visible. These photoluminescent carbon dots (C-Dots, Figure 1a) are of two distinctive features: one is that the underlying carbon particles are very small (sub-10 nm); and the other is that the particle surface is passivated by organic or other molecules via either covalent linkages or chemical adsorption. 11 Mechanistically, the carbon-based photoluminescence has been attributed to passivated defects on the carbon particle surface acting as excitation energy traps. 11,12 Here we report that the C-Dots also exhibit strong luminescence with two-photon excitation in the near-infrared. The estimated two-photon absorption cross-sections of the C-Dots are comparable to those of available high-performance semiconductor quantum dots. In addition, the two-photon luminescence microscopy imaging of the C-Dots internalized in human cancer cells is demonstrated.
There has been significant recent interest in the development of highly fluorescent nanomaterials as contrast agents for optical imaging in vivo. 1 The imaging agents should ideally be bright, nontoxic, biocompatible, and stable against photobleaching. Among the extensively studied are those based on semiconductor quantum dots (QDs) such as CdSe/ZnS. 2 The rationale for the use of QDs over conventional organic dyes is now generally accepted in the literature. 3 There are already successful in vivo imaging demonstrations of QDs on tumor vasculature, tumor-specific membrane antigens, sentinel lymph nodes, etc. 2,4 The semiconductor QDs containing cadmium or other heavy metals are unfortunately known for their significant toxicity even at relatively low concentrations, 5,6 which may prove prohibitive to any patient studies. Therefore, the search for benign alternatives has continued. Of particular interest and significance was the recent finding that small carbon nanoparticles could be surface-passivated by organic or bio-molecules to become strongly fluorescent. 7 These fluorescent carbon nanoparticles, 7,8 dubbed "carbon dots" (C-Dots, Scheme 1), were found to be physicochemically and photochemically stable and non-blinking. The carbon particle core could also be doped with an inorganic salt such as ZnS before the surface functionalization to significantly enhance the fluorescence brightness (C ZnS -Dots, Scheme 1). 9 These carbon dots have been successfully used for in vitro cell imaging with both one-and two-photon excitations. 7,9,10 Carbon is hardly considered as an intrinsically toxic element. Available results from the ongoing toxicity evaluation of the oligomeric PEG-functionalized C-Dots 7 in mice have suggested no meaningful toxic effects, 11 raising the prospect for in vivo biocompatibility and uses of carbon dots. Here we report the first study of carbon dots for optical imaging in vivo. The results suggest that the carbon dots are not only brightly fluorescent in solution, as reported previously, 7,9 but also well-behaved as contrast agents in live mice.The C-Dots and C ZnS -Dots with the PEG diamine, H 2 NCH 2 (CH 2 CH 2 O) n CH 2 CH 2 CH 2 NH 2 (n ∼ 35, PEG 1500N ), as the surface passivation agent were prepared and characterized as previously reported. 7,9,10 Shown in Figure 1 For subcutaneous injection, female DBA/1 mice (∼25 g) were shaved in the back area surrounding the injection point. Upon the injection of a C-Dots solution (30 µg carbon coreequivalent in 30 µL) or a C ZnS -Dots solution (65 µg in 30 µL), the mice were imagined in a Lumazone FA in vivo Imaging System (MAG Biosystems) with 470 nm (FWHM ∼ 40 nm) excitation and 525 nm (FWHM ∼ 47 nm) emission filters. As shown in Figure 2, the fluorescence images of the subcutaneously injected mice exhibited bright emissions from CDots and C ZnS -Dots. The relatively stronger fluorescence from the latter is consistent with the previously reported solution-phase results. 9 The injected carbon dots in mice diffused relatively slowly, with the ...
Fluorescent carbon dots (small carbon nanoparticles with the surface passivated by oligomeric PEG molecules) were evaluated for their cytotoxicity and in vivo toxicity and also for their optical imaging performance in reference to that of the commercially supplied CdSe/ZnS quantum dots. The results suggested that the carbon dots were biocompatible, and their performance as fluorescence imaging agents was competitive. The implication to the use of carbon dots for in vitro and in vivo applications is discussed.
There have been multiple conflicting reports about the biocompatibility and antimicrobial activity of graphene oxide. To address this, we conducted a study to characterize the antimicrobial properties of graphene oxide (GO) and its biocompatibility with mammalian cells. When GO was added to a bacterial culture at 25 μg/mL, the results showed that bacteria grew faster and to a higher optical density than cultures without GO. Scanning electron microscopy indicated that bacteria formed dense biofilms in the presence of GO. This was shown by a large mass of aggregated cells and extracellular polymeric material. Bacterial growth on filters coated with 25 and 75 μg of GO grew 2 and 3 times better than on filters without GO. Closer analysis showed that bacteria were able to attach and proliferate preferentially in areas containing the highest GO levels. Graphene oxide films failed to produce growth inhibition zones around them, indicating a lack of antibacterial properties. Also, bacteria were able to grow on GO films to 9.5 × 10(9) cells from an initial inoculation of 1.0 × 10(6), indicating that it also lacks bacteriostatic activity. Thus, silver-coated GO films were able to produce clearing zones and cell death. Also, graphene oxide was shown to greatly enhance the attachment and proliferation of mammalian cells. This study conclusively demonstrates that graphene oxide does not have intrinsic antibacterial, bacteriostatic, and cytotoxic properties in both bacteria and mammalian cells. Furthermore, graphene oxide acts as a general enhancer of cellular growth by increasing cell attachment and proliferation.
Carbon dots (or carbon quantum dots in some literature reports), generally small carbon nanoparticles with various surface passivation effects, have attracted widespread attention in recent years, with a rapidly increasing number of research publications. The reported studies covered many aspects of carbon dots, from the development of many new synthetic methodologies to an improved mechanistic elucidation and to the exploration of application opportunities, especially for those in the fluorescence imaging of cells and tissues. There have also been significant advances in the establishment of a shared mechanistic framework for carbon dots and other carbon-based quantum dots, graphene quantum dots in particular.In this article, representative recent studies for more efficient syntheses of better-performing carbon dots are highlighted along with results from explorations of their various bioimaging applications in vitro and in vivo. Similar fluorescence properties and potential imaging uses of some graphene quantum dots are also discussed, toward a more consistent and uniform understanding of phenomenologically different carbon-based quantum dots. Pengju G. Luo has been a faculty member atSherman College of Chiropractic since 2006. He received his medical degree in Clinical Medicine from Tongji Medical University, Wuhan, China in 1997 and his Ph.D. in Microbiology from Clemson University in 2006. His current research interests are in the interdisciplinary areas of nanotechnology and biological and biomedical sciences, focusing on bioapplications of various nanomaterials such as nanoparticles, nanotubes, and carbon dots. Fan Yang obtained his B.S. degree in Chemistry from Zhejiang University, China in 2010. He is currently pursuing his Ph.D. at Clemson University under the supervision of Prof. Ya-Ping Sun. His research focus is on the synthesis of carbon dots for their applications in bioimaging and energy conversions.
This progress report provides an overview on recent advances in bioapplications of carbon nanotubes including the chemical modification of carbon nanotubes, targeting specifically their covalent and noncovalent conjugations with a variety of biological and bioactive species (proteins and peptides, DNAs/RNAs, and carbohydrates). Furthermore, the significant recent development and progress in the use of carbon nanotubes for biosensors, drug and other delivery systems, bioimaging, etc. and in the understanding of in vivo biodistribution and toxicity of carbon nanotubes are reported.
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