Nature-Inspired Stalactite Nanopores for Biosensing and Energy Harvesting.

Nature provides a wide range of self-assembled structures from the nanoscale to the macroscale. Under the right thermodynamic conditions and with the appropriate material supply, structures like stalactites, icicles, and corals can grow. However, the natural growth process is time-consuming. This work demonstrates a fast, nature-inspired method for growing stalactite nanopores using heterogeneous atomic deposition of hafnium dioxide at the orifice of templated silicon nitride apertures. The stalactite nanostructures combine the benefits of reduced sensing region typically for 2-dimensional material nanopores with the asymmetric geometry of capillaries, resulting in ionic selectivity, stability, and scalability. The proposed growing method provides an adaptable nanopore platform for basic and applied nanofluidic research, including biosensing, energy science, and filtration technologies.

High durability and stability of 2D nanofluidic devices for long-term single-molecule sensing.

Nanopores in two-dimensional (2D) membranes hold immense potential in single-molecule sensing, osmotic power generation, and information storage. Recent advances in 2D nanopores, especially on single-layer MoS2, focus on the scalable growth and manufacturing of nanopore devices. However, there still remains a bottleneck in controlling the nanopore stability in atomically thin membranes. Here, we evaluate the major factors responsible for the instability of the monolayer MoS2 nanopores. We identify chemical oxidation and delamination of monolayers from their underlying substrates as the major reasons for the instability of MoS2 nanopores. Surface modification of the substrate and reducing the oxygen from the measurement solution improves nanopore stability and dramatically increases their shelf-life. Understanding nanopore growth and stability can provide insights into controlling the pore size, shape and can enable long-term measurements with a high signal-to-noise ratio and engineering durable nanopore devices.

Wafer-scale MoS2 with water-vapor assisted showerhead MOCVD.

Among numerous thin film synthesis methods, metalorganic chemical vapor deposition performed in a showerhead reactor is the most promising one for broad use in scalable and commercially adaptable two-dimensional material synthesis processes. Adapting the most efficient monolayer growth methodologies from tube-furnace systems to vertical-showerhead geometries allows us to overcome the intrinsic process limitations and improve the overall monolayer yield quality. Here, we demonstrate large-area, monolayer molybdenum disulphide growth by combining gas-phase precursor supply with unique tube-furnace approaches of utilizing sodium molybdate pre-seeding solution spincoated on a substrate along with water vapor added during the growth step. The engineered process yields a high-quality, 4-inch scale monolayer film on sapphire wafers. The monolayer growth coverage, average crystal size and defect density were evaluated using Raman and photoluminescence spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscopy and scanning transmission electron microscopy imaging. Our findings provide a direct step forward toward developing a reproducible and large-scale MoS2 synthesis with commercial showerhead reactors.

High-Throughput Nanopore Fabrication and Classification Using Xe-Ion Irradiation and Automated Pore-Edge Analysis.

Large-area nanopore drilling is a major bottleneck in state-of-the-art nanoporous 2D membrane fabrication protocols. In addition, high-quality structural and statistical descriptions of as-fabricated porous membranes are key to predicting the corresponding membrane-wide permeation properties. In this work, we investigate Xe-ion focused ion beam as a tool for scalable, large-area nanopore fabrication on atomically thin, free-standing molybdenum disulfide. The presented irradiation protocol enables designing ultrathin membranes with tunable porosity and pore dimensions, along with spatial uniformity across large-area substrates. Fabricated nanoporous membranes are then characterized using scanning transmission electron microscopy imaging, and the observed nanopore geometries are analyzed through a pore-edge detection and analysis script. We further demonstrate that the obtained structural and statistical data can be readily passed on to computational and analytical tools to predict the permeation properties at both individual pore and membrane-wide scales. As an example, membranes featuring angstrom-scale pores are investigated in terms of their emerging water and ion flow properties through extensive all-atom molecular dynamics simulations. We believe that the combination of experimental and analytical approaches presented here will yield accurate physics-based property estimates and thus potentially enable a true function-by-design approach to fabrication for applications such as osmotic power generation and desalination/filtration.

Stress induced delamination of suspended MoS2 in aqueous environments.

Applying hydrostatic pressure with suspended 2D material thin membranes allows probing the effects of lateral strain on the ion and fluid transport through nanopores. We demonstrate how both permanent and temporary delamination of 2D materials can be induced by pressure and potential differences between the membrane, causing a strong mechanosensitive modulation of ion transport. Our methodology is based on in situ measurements of ionic current and streaming modulation as the supporting membrane is indented or bulged with pressure. We demonstrate how indirect measurements of fluid transport through delaminated MoS2 membranes is achieved through monitoring streaming current and potential. This is combined with TEM images of 2D material membranes before and after aqueous measurements, showing temporary delamination during mechanical or electrical stress. The obtained results allow a better understanding of measurements with supported 2D materials, i.e. avoiding misinterpreting measured data, and could be used to probe how the electrical field and fluid flow at the nanoscale influence the adhesion of supported 2D materials.

Preparation of graphene oxide-graphene quantum dots hybrid and its application in cancer theranostics.

Currently, an enormous amount of cancer research based on two-dimensional nano-graphene oxide (GO), as well as zero-dimensional graphene quantum dots (GQDs), is being carried out in the fields of therapeutics and diagnostics. However, the exploration of their hybrid "functional" nanomaterials in the theranostic system is still rare. In the current study, a stable complex of GO and GQDs was formed by an electrostatic layer-by-layer assembly via a polyethylene imine bridge (GO-PEI-GQDs). Furthermore, we compared separate mono-equivalents of the GO-PEI-GQDs complex - GO and GQDs, in terms of cell imaging (diagnostics), photothermal, and oxidative stress response in breast cancer cells (MDA-MB-231). GO-PEI-GQDs showed an excellent photothermal response (44-49 °C) upon 808 nm laser (0.5 W cm-2) exposure for 5 min at a concentration up to 50 µg/mL. We report new synergistic properties of GO-PEI-GQDs such as stable fluorescence imaging and enhanced photothermal and cytotoxic activities on cancer cells. Composite materials made up of GO and GQDs combining diverse properties help to study 2D-0D heterosystems and improve specific therapeutic systems in theranostics.

Fabrication and practical applications of molybdenum disulfide nanopores.

Among the different developed solid-state nanopores, nanopores constructed in a monolayer of molybdenum disulfide (MoS2) stand out as powerful devices for single-molecule analysis or osmotic power generation. Because the ionic current through a nanopore is inversely proportional to the thickness of the pore, ultrathin membranes have the advantage of providing relatively high ionic currents at very small pore sizes. This increases the signal generated during translocation of biomolecules and improves the nanopores' efficiency when used for desalination or reverse electrodialysis applications. The atomic thickness of MoS2 nanopores approaches the inter-base distance of DNA, creating a potential candidate for DNA sequencing. In terms of geometry, MoS2 nanopores have a well-defined vertical profile due to their atomic thickness, which eliminates any unwanted effects associated with uneven pore profiles observed in other materials. This protocol details all the necessary procedures for the fabrication of solid-state devices. We discuss different methods for transfer of monolayer MoS2, different approaches for the creation of nanopores, their applicability in detecting DNA translocations and the analysis of translocation data through open-source programming packages. We present anticipated results through the application of our nanopores in DNA translocations and osmotic power generation. The procedure comprises four parts: fabrication of devices (2-3 d), transfer of MoS2 and cleaning procedure (24 h), the creation of nanopores within MoS2 (30 min) and performing DNA translocations (2-3 h). We anticipate that our protocol will enable large-scale manufacturing of single-molecule-analysis devices as well as next-generation DNA sequencing.

Graphene Quantum Dots for Cell Proliferation, Nucleus Imaging, and Photoluminescent Sensing Applications.

We report a simple one-pot microwave assisted "green synthesis" of Graphene Quantum Dots (GQDs) using grape seed extract as a green therapeutic carbon source. These GQDs readily self-assemble, hereafter referred to as "self-assembled" GQDs (sGQDs) in the aqueous medium. The sGQDs enter via caveolae and clathrin-mediated endocytosis and target themselves into cell nucleus within 6-8 h without additional assistance of external capping/targeting agent. The tendency to self-localize themselves into cell nucleus also remains consistent in different cell lines such as L929, HT-1080, MIA PaCa-2, HeLa, and MG-63 cells, thereby serving as a nucleus labelling agent. Furthermore, the sGQDs are highly biocompatible and act as an enhancer in cell proliferation in mouse fibroblasts as confirmed by in vitro wound scratch assay and cell cycle analysis. Also, photoluminescence property of sGQDs (lifetime circa (ca.) 10 ns) was used for optical pH sensing application. The sGQDs show linear, cyclic and reversible trend in its fluorescence intensity between pH 3 and pH 10 (response time: ~1 min, sensitivity -49.96 ± 3.5 mV/pH) thereby serving as a good pH sensing agent. A simple, cost-effective, scalable and green synthetic approach based sGQDs can be used to develop selective organelle labelling, nucleus targeting in theranostics, and optical sensing probes.

Evolution of thiol-capped gold nanoclusters into larger gold nanoparticles under electron beam irradiation.

We report in situ transformation of glutathione-capped red-fluorescent gold nanoclusters (AuNCs) into larger gold nanoparticles (AuNPs) embedded on a copper grid under high energy electron beam of Field Emission Gun Transmission Electron Microscope (FEG-TEM). Electron beam irradiation causes coalescing of individual finer AuNCs into bigger discrete AuNPs as a function of electron dose rate and time. The coalescence was closely studied over time and the mechanism is discussed. The study will help to understand the structural and morphological changes that occur in AuNCs inside FEG-TEM due to prolonged electron beam exposure.

Milk-derived multi-fluorescent graphene quantum dot-based cancer theranostic system.

An economical green-chemistry approach was used for the synthesis of aqueous soluble graphene quantum dots (GQDs) from cow milk for simultaneous imaging and drug delivery in cancer. The GQDs synthesized using one-pot microwave-assisted heating were multi-fluorescent, spherical in shape having a lateral size of ca. 5nm. The role of processing parameters such as heating time and ionic strength showed a profound effect on photoluminescence properties of GQDs. The GQDs were N-doped and oxygen-rich as confirmed by X-ray photoelectron spectroscopy (XPS) analysis. Cysteamine hydrochloride (Cys) was used to attach an anti-cancer drug berberine hydrochloride (BHC) on GQDs forming GQDs@Cys-BHC complex with c.a. 88% drug loading efficiency. In vitro drug release was studied at the acidic-basic environment and drug kinetics was studied using pharmacokinetic statistical models. The GQDs were biocompatible on L929 cells whereas theranostic GQDs@Cys-BHC complex showed a potent cytotoxic effect on different cancerous cell line models: cervical cancer cell lines such as HeLa cells and breast cancer cells such as MDA-MB-231 confirmed by Trypan blue and MTT-based cytotoxic assays. Furthermore, multi-excitation based cellular bioimaging was demonstrated using confocal laser scanning microscopy (CLSM) and fluorescence microscopy using GQDs as well as GQDs@Cys-BHC complex. Thus, drug delivery (therapeutic) and bioimaging (diagnostic) properties of GQDs@Cys-BHC complex are thought to have a potential in vitro theranostic application in cancer therapy.

Biogenic Synthesis of Fluorescent Carbon Dots at Ambient Temperature Using Azadirachta indica (Neem) gum.

Synthesis of fluorescent Carbon Dots (CDs) from various carbonaceous materials apparently has acquired lots of interest amongst researchers as the corollary of the properties of CDs; which are subsequently getting unveiled. In this study we report the use of Azadirachta indica (Neem) Gum as a novel natural pre-cursor for synthesis of CDs at room temperature. Water soluble CDs of around 5-8 nm were obtained after treatment of the gum with ethanol and NaOH. These CDs exhibited green fluorescence in UV-light (λ = 365 nm). These CDs were found to be stable, having many bio-linkers attached on their surface, making it suitable for drug attachment and hence can serve as potential candidates for applications like drug delivery vehicles as well as for biosensors.

Synthesis of highly fluorescent hydrophobic carbon dots by hot injection method using Paraplast as precursor.

We have reported synthesis of bright blue colored hydrophobic carbon dots (hC-dots) using highly pure blend of polymers called Paraplast. We developed a hot injection method for making nearly monodispersed hC-dots with a diameter in a range: 5-30nm as confirmed by high resolution transmission electron microscopy (HRTEM). The involvement of various functional groups was confirmed by Fourier transform infra-red (FTIR) spectroscopy. These hC-dots were incubated with breast cancer stem cells in order to check the entry as well as biological imaging. The cells were analyzed using epifluorescent microscopy. hC-dots showed concentration dependent cytotoxicity (LD50: 50mg/ml) and could be used for bioimaging even at lower concentration (0.5mg/ml). hC-dots were found to be versatile agents for peeping inside the cells which could also be used for delivery of water insoluble chemotherapeutic agents to variety of solid tumors.

A green route towards highly photoluminescent and cytocompatible carbon dot synthesis and its separation using sucrose density gradient centrifugation.

An efficient, fast and green method for synthesis of Carbon dots (C-dots) using natural precursor Citrus limone under ultrasonic condition is demonstrated. Such as-synthesized C-dots were further purified using Sucrose density gradient centrifugation method (SDGC) which resulted in the separation of water-soluble, photo luminescent, monodispersed, highly photostable and chemically stable C-dot fractions (F1 and F2). They possess very small size (5-20 nm) as evidenced by High angle annular dark field-Scanning Transmission electron microscopy (HAADF-STEM) and very strong luminescence as shown by fluorescence spectroscopic studies. Cytocompatibility and bio imaging properties of both the fractions (F1 and F2) were then studied on Hep-2 cells. Quantum yield of F1 and F2 fraction was found to be 12.1 and 15 %, respectively.

Biogenic gold nano-triangles: cargos for anticancer drug delivery.

We present synthesis of biogenic gold nano triangles (GNTs) using Azadirachta indica leaf extract at inherent pH (5.89) and its application in efficient drug delivery of doxorubicin (DOX) (anticancer drug). The main idea was to take advantage of large surface area of GNTs which has 3 dimensions and use the plant peptides coated on these triangles as natural linkers for the attachment of DOX. Sucrose density gradient centrifugation (SDGC) and dialysis methods were used for separation of the GNT from mixture of GNPs. Flocculation parameter (FP) was used to check stability of GNT which was found to be exceptionally high (0-0.75) due to the biological capping agents. DOX attachment to GNT was verified using Fourier transformed infra-red (FTIR) spectroscopy. The complex thus formed was found to be less toxic to normal cells (MDCK cells) and significantly toxic for the cancerous cells (HeLa cells). Drug loading efficiency was found to be 99.81% and DOX release followed first order release kinetics. Percentage drug release was found to be more than 4.5% in both acidic (5.8) as well as physiological pH (7.2) which is suitable for tumor targeting.

Antibiotic conjugated fluorescent carbon dots as a theranostic agent for controlled drug release, bioimaging, and enhanced antimicrobial activity.

A novel report on microwave assisted synthesis of bright carbon dots (C-dots) using gum arabic (GA) and its use as molecular vehicle to ferry ciprofloxacin hydrochloride, a broad spectrum antibiotic, is reported in the present work. Density gradient centrifugation (DGC) was used to separate different types of C-dots. After careful analysis of the fractions obtained after centrifugation, ciprofloxacin was attached to synthesize ciprofloxacin conjugated with C-dots (Cipro @ C-dots conjugate). Release of ciprofloxacin was found to be extremely regulated under physiological conditions. Cipro @ C-dots were found to be biocompatible on Vero cells as compared to free ciprofloxacin (1.2 mM) even at very high concentrations. Bare C-dots ( ∼ 13 mg mL(-1)) were used for microbial imaging of the simplest eukaryotic model-Saccharomyces cerevisiae (yeast). Bright green fluorescent was obtained when live imaging was performed to view yeast cells under fluorescent microscope suggesting C-dots incorporation inside the cells. Cipro @ C-dots conjugate also showed enhanced antimicrobial activity against both model gram positive and gram negative microorganisms. Thus, the Cipro @ C-dots conjugate paves not only a way for bioimaging but also an efficient new nanocarrier for controlled drug release with high antimicrobial activity, thereby serving potential tool for theranostics.

Swarming carbon dots for folic acid mediated delivery of doxorubicin and biological imaging.

Carbon dots (C-dots) are one of the most highlighted carbon-based materials for biological applications such as delivery of therapeutic payloads for cancer treatment mainly due to their biocompatibility and unique optical properties. In this work, we have explored the drug carrying capacity of highly fluorescent sorbitol-derived C-dots for targeted delivery of doxorubicin (DOX). We have used folic acid (FA) as a navigational molecule due to its high expression in most cancer cells. Before attachment of the DOX, the surfaces of the C-dots were protected with bovine serum albumin (BSA) to make them more biocompatible and able to hold a high amount of drugs. The release profile of DOX was studied using standard statistical models and confirmed to be first order at pH 7.2. Cellular imaging was performed using epifluorescence microscopy, which showed bright green coloured fluorescence due to internalization of C-dots specifically targeted with FA in HeLa cells.

Biogenic gold nanoparticles as fotillas to fire berberine hydrochloride using folic acid as molecular road map.

Use of biologically modified gold nanoparticles (GNPs) as molecular vehicle to ferry potential anti-cancer drug berberine hydrochloride (BHC) using folic acid (FA) as targeting molecule is reported in this work. A tropical fruit peel, Trapa bispinosa is used to fabricate highly monodispersed GNPs, passivated with essential functional groups which were used as linkers to attach FA and BHC via amide linkage. Flocculation Parameter (FP) of biologically synthesized GNPs was calculated under different salt concentrations which were found to be very ideal under a physiological condition. Various statistical models were used to find drug release profile out of which Higuchi was found to be the most ideal. GNP-FA-BHC complexes were found to be active against folic acid expressing HeLa cells.

A comparative study of economical separation and aggregation properties of biologically capped and thiol functionalized gold nanoparticles: selecting the eco-friendly trojan horses for biological applications.

We are presenting facile bio-fabrication of extremely stable gold nanoparticles (GNPs) using medicinal plant Azadirachta indica (commonly called Neem) and its comparison with most commonly used glutathione (GSH) protected GNPs in terms of stability under physiological conditions, seperation using density gradient centrifugation and aggregation properties in the solution. There was dual peak at 536 and 662 nm indicating the presence of non-spherical GNPs including triangles, rods and hexagons in case of A. indica mediated GNPs unlike citrate stabilized GNPs which exhibited single sharp peak. Spherical GNPs were separated from the consortium of uniquely shaped nanoparticles bio-fabricated using A. indica leaf extract using sucrose density gradient centrifugation (SDGC).To comprehend the anti-agglomeration potentials of A. indica leaf mediated GNPs and GSH-GNPs under physiological conditions, flocculation parameters (FP) were calculated and found to be least for A. indica leaf mediated GNPs, indicating their exceptional stability.

Green synthesis of biocompatible carbon dots using aqueous extract of Trapa bispinosa peel.

We are reporting highly economical plant based method for the production of luminescent water soluble carbon dots (C-dot) using Indian water plant Trapa bispinosa peel extract without adding any external oxidizing agent at 90 °C. C-dots ranging from 5 to 10nm were found in the solution with a prominent green fluorescence under UV-light (λex=365 nm). UV-vis spectra recorded at different time intervals (30-120 min) displayed signature absorption of C-dots between 400 and 600 nm. Fluorescence spectra of the dispersion after 120 min of synthesis exhibited characteristic emission peaks of C-dots when excited at 350, 400, 450 and 500 nm. C-dots were further analyzed using X-ray diffraction (XRD), Raman Spectroscopy and Thermo-Gravimetric Analysis (TGA). Structure of the C-dots was found to be turbostratic when studied using XRD. C-dots synthesized by our method were found to be exceptionally biocompatible against MDCK cells.

Understanding the stability of silver nanoparticles bio-fabricated using Acacia arabica (Babool gum) and its hostile effect on microorganisms.

We report green synthesis of stable silver nanoparticles (SNPs) from Acacia arabica gum and its anti-bacterial activity against gram-positive and gram-negative bacteria. UV-Vis spectral analysis of synthesized SNPs showed maximum peak at 462 nm initially and 435 nm after 24 h. Using Transmission Electron microscopy (TEM), the average size of synthesised SNPs was found to be ~35 nm. X-ray diffraction (XRD) and Selective area electron diffraction (SAED) pattern confirmed the crystalline nature of SNPs. Percentage conversion of Ag(+) ions into Ag° was calculated using ICP-AES and was found to be 94%. By calculating flocculation parameter, we could see that these SNPs are extremely stable under the influence of very high NaCl concentration up to 4.16 M. These stable SNPs can be used in various industrial and medical applications.