Gold nanoshell photomodification under a single-nanosecond laser pulse accompanied by color-shifting and bubble formation phenomena

Laser-nanoparticle interaction is crucial for biomedical applications of lasers and nanotechnology to the treatment of cancer or pathogenic microorganisms. We report on the first observation of laser-induced coloring of gold nanoshell solution after a one nanosecond pulse and an unprecedentedly low bubble formation (as the main mechanism of cancer cell killing) threshold at a laser fluence of about 4?mJ?cm(-2), which is safe for normal tissue. Specifically, silica/gold nanoshell (140/15?nm) suspensions were irradiated with a single 4?ns (1064?nm) or 8?ns (900?nm) laser pulse at fluences ranging from 0.1?mJ?cm(-2) to 50?J?cm(-2). Solution red coloring was observed by the naked eye confirmed by blue-shifting of the absorption spectrum maximum from the initial 900?nm for nanoshells to 530?nm for conventional colloidal gold nanospheres. TEM images revealed significant photomodification of nanoparticles including complete fragmentation of gold shells, changes in silica core structure, formation of small 20-30?nm isolated spherical gold nanoparticles, gold nanoshells with central holes, and large and small spherical gold particles attached to a silica core. The time-resolved monitoring of bubble formation phenomena with the photothermal (PT) thermolens technique demonstrated that after application of a single 8?ns pulse at fluences 5-10?mJ?cm(-2) and higher the next pulse did not produce any PT response, indicating a dramatic decrease in absorption because of gold shell modification. We also observed a dependence of the bubble expansion time on the laser energy with unusually very fast PT signal rising (～3.5?ns scale at 0.2?J?cm(-2)). Application of the observed phenomena to medical applications is discussed, including a simple visual color test for laser-nanoparticle interaction.     

Evaluation of red CdTe and near infrared CdHgTe quantum dots by fluorescent imaging

Non-invasive fluorescent imaging of preclinical animal models in vivo is a rapidly developing field with new emerging technologies and techniques. Quantum dot (QD) fluorescent probes with longer emission wavelengths in red and near infrared (NIR) emission ranges are more amenable to deep-tissue imaging, because both scattering and autofluorescence are reduced as wavelengths are increased. We have designed and synthesized red CdTe and NIR CdHgTe QDs for fluorescent imaging. We demonstrated fluorescent imaging by using CdTe and CdHgTe QDs as fluorescent probes both in vitro and in vivo. Both CdTe and CdHgTe QDs provided sensitive detection over background autofluorescence in tissue biopsies and live mice, making them attractive probes for in vivo imaging extending into deep tissues or whole animals. The studies suggest a basis of using QD-antibody conjugates to detect membrane antigens.     

Minimizing nonspecific cellular binding of quantum dots with hydroxyl-derivatized surface coatings

Quantum-dot (QD) nanocrystals are promising fluorescent probes for multiplexed staining assays in biological applications. However, nonspecific QD binding to cellular membranes and proteins remains a limiting factor in detection sensitivity and specificity. Here we report a new class of hydroxyl (-OH)-coated QDs for minimizing nonspecific cellular binding and for overcoming the bulky size problems encountered with previous surface coatings. The hydroxylated QDs are prepared from carboxylated (-COOH) dots via a hydroxylation and cross-linking process. With a compact hydrodynamic size of 13-14 nm (diameter), they are highly fluorescent (>60% quantum yields) and stable under both basic and acidic conditions. By using human cancer cells, we have evaluated their superior nonspecific binding properties against that of carboxylated, protein-coated, and poly(ethylene glycol) (PEG)-coated QDs. Quantitative cellular staining data indicate that the hydroxylated QDs result in a dramatic 140-fold reduction in nonspecific binding relative to that of carboxylated dots and a still significant 10-20-fold reduction relative to that of PEG- and protein-coated dots.     

Aqueous CdPbS quantum dots for near-infrared imaging

Quantum dots (QDs) are semiconducting nanocrystals that have photoluminescent (PL) properties brighter than fluorescent molecules and do not photo-bleach, ideal for in vivo imaging of diseased tissues or monitoring of biological processes. Near-infrared (NIR) fluorescent light within the window of 700-1000 nm, which is separated from the major absorption peaks of hemoglobin and water, has the potential to be detected several millimeters under the surface with minimal interference from tissue autofluorescence. Here we report the synthesis and bioimaging demonstration of a new NIR QDs system, namely, CdPbS, made by an aqueous approach with 3-mercaptopropionic acid (MPA) as the capping molecule. The aqueous-synthesized, MPA-capped CdPbS QDs exhibited an NIR emission in the range of 800-950 nm with x(i) ≥ 0.3, where x(i) denotes the initial Pb molar fraction during the synthesis. Optimal PL performance of the CdPbS QDs occurred at x(i) = 0.7, which was about 4 nm in size as determined by transmission electron microscopy, had a rock salt structure and a quantum yield of 12%. Imaging of CdPbS QDs was tested in membrane staining and transfection studies. Cells transfected with CdPbS QDs were shown to be visible underneath a slab of chicken muscle tissue of up to 0.7 mm in thickness without the use of multiple-photon microscopy.     

Semiconductor Quantum Dots for Biosensing and In Vivo Imaging

Semiconductor quantum dots (QDs) have captivated researchers in the biomedical field over the last decade. Compared to organic dyes and fluorescent proteins, QDs have unique optical properties such as tunable emission spectra, improved brightness, superior photostability, and simultaneous excitation of multiple fluorescence colors. Since the first successful reports on the biological use of QDs a decade ago, QDs and their bioconjugates have been successfully applied to various imaging applications including fixed cell labeling, live-cell imaging, in situ tissue profiling, fluorescence detection and sensing, and in vivo animal imaging. In this review, we will briefly survey the optical properties of QDs, the biofunctionalization strategies, and focus on their biosensing and in vivo imaging applications. We conclude with a discussion on the issues and perspectives on QDs as biosensing probes and in vivo imaging agents.      

One-pot aqueous synthesis PbS quantum dots and their Hg2+ sensitive properties

In this study, we report the synthesis and stability of PbS quantum dots (QDs) using an aqueous route with L-Cysteine (L-Cys) as the capping molecule. The as-synthesized L-Cys-capped PbS QDs were characterized by high resolution transmission electron microscopy (HRTEM), and X-ray diffraction (XRD), the results indicated that the QDs were about 4 nm in size and dispersed well with a rock salt crystalline structure, and there was L-Cys on the surface of QDs, which was confirmed by Fourier transform infrared (FT-IR) spectrometry. The influence of various experimental variables, including amounts of capping ligand, pH value and refluxing time, on the luminescent properties of the obtained QDs have been systematically investigated. The QDs exhibited optimal PL intensity when Pb: L-Cys: S = 1:2.2:0.3. In addition, the as-prepared QDs could be used as fluorescence probes to detect Hg2+ ions in aqueous media. The response of QDs fluorescence probes was linearly proportional to the concentration of Hg2+ ions ranging from 8 x 10(-9) to 2 x 10(-6) mol x L(-1) with a limit of detection of 2 x 10(-9) mol x L(-1). Furthermore, the method was successfully applied to the determine Hg2+ ions in different real samples.     

Fluorescent magnetic hybrid nanoprobe for multimodal bioimaging

A fluorescent magnetic hybrid imaging nanoprobe (HINP) was fabricated by the conjugation of superparamagnetic Fe3O4 nanoparticles and visible light emitting (～600 nm) fluorescent CdTe/CdS quantum dots (QDs). The assembly strategy used the covalent linking of the oxidized dextran shell of magnetic particles to the glutathione ligands of QDs. The synthesized HINP formed stable water-soluble colloidal dispersions. The structure and properties of the particles were characterized by transmission electron and atomic force microscopy, energy dispersive x-ray analysis and inductively coupled plasma optical emission spectroscopy, dynamic light scattering analysis, optical absorption and photoluminescence spectroscopy, and fluorescent imaging. The luminescence imaging region of the nanoprobe was extended to the near-infrared (NIR) (～800 nm) by conjugation of the superparamagnetic nanoparticles with synthesized CdHgTe/CdS QDs. Cadmium, mercury based QDs in HINP can be easily replaced by novel water-soluble glutathione stabilized AgInS2/ZnS QDs to present a new class of cadmium-free multimodal imaging agents. The observed NIR photoluminescence of fluorescent magnetic nanocomposites supports their use for bioimaging. The developed HINP provides dual-imaging channels for simultaneous optical and magnetic resonance imaging.     

Exploring feasibility of multicolored CdTe quantum dots for in vitro and in vivo fluorescent imaging

We report the use of novel multicolored CdTe quantum dots (QDs) as fluorophores for biological fluorescence imaging. The CdTe QDs were prepared to exhibit emission wavelengths in the green, yellow, and red range by using trifluoroacetic acid (TFA), L-cysteine and thioglycolic acid (TGA) as surface stabilizers, respectively. The particles have good water solubility and photostability. Fluorescence imaging potential was evaluated in vitro and in vivo using a multispectral Maestro CRI Fluorescence Imaging system. The results show that different colored CdTe QDs allow sensitive detection simultaneously or separately both in vitro and in vivo against background fluorescence. The studies indicate that CdTe QDs can provide alternative fluorescent probes for biological imaging.     

Hybrid confocal Raman fluorescence microscopy on single cells using semiconductor quantum dots

We have overcome the traditional incompatibility of Raman microscopy with fluorescence microscopy by exploiting the optical properties of semiconductor fluorescent quantum dots (QDs). Here we present a hybrid Raman fluorescence spectral imaging approach for single-cell microscopy applications. We show that resonant Raman imaging of flavocytochrome b558 at 413.1 nm excitation in QD-labeled neutrophilic granulocytes or nonresonant Raman imaging of proteins and lipids at 647.1 nm excitation in QD-labeled macrophages can be integrated with linear one-photon excitation and nonlinear continuous-wave two-photon excitation fluorescence microscopy of QDs, respectively. The enhanced information content of these two hybrid Raman fluorescence methods provides new multiplexing possibilities for single-cell optical microscopy and intracellular chemical analysis.     

Quantum dot conjugated hydroxylapatite nanoparticles for in vivo imaging

Hydroxylapatite (HA) nanoparticles were conjugated with quantum dots (QDs) for in vivo imaging. The surface structures of HA nanoparticles with conjugated quantum dots (HA-QD) were studied by transmission electron microscopy (TEM) and laser fluorescent spectroscopy. The TEM data showed that the quantum dots were well conjugated on the HA nanoparticle surfaces. The laser fluorescent spectroscopy results indicated that the HA-QD exhibited promising luminescent emission in vitro. The initial in vivo experiments revealed clear images of HA-QD from the hypodermic injected area at the emission of 600?nm. Furthermore, the optimized in vivo images of HA-QD with near-infrared emission at 800?nm were visualized after intravenous injection. These luminescent HA-QD nanoparticles may find important applications as biodegradable substrates for biomarkers and in drug delivery.     

近红外荧光磁性复合载药脂质体的制备及应用

拟建立以近红外荧光磁性复合脂质体(NFMSLs)为模型药物载体、盐酸多柔比星(DOX)为包封药物的药物输送系统,研究了近红外荧光磁性载药复合脂质体(DOX-NFMSLs)的制备、性质及初步应用.采用共沉淀法制备FeO4磁流体,CdTe掺杂Se制备近红外量子点CdSeTe,薄膜分散法制备DOX-NFMSLs.用DOX荧光分光光度法测定DOX-NFMSLs的包封率和体外药物释放率;用DOX-NFMSLs与HepG2肝癌细胞共孵育来进行细胞成像和细胞毒性实验.结果表明,近红外CdSeTe量子点粒径约为5nm,闪锌矿结构,发射波长824 nm.磷脂与胆固醇质量比为8∶1,药脂比为1∶20的DOX-NFMSLs平均粒径为252.9 nm,Zeta电位为-48.6 mV,理想释放药物温度为41℃,平均包封率为(74.84±0.89)％.DOX-NFMSLs对HepG2肝癌细胞有一定的抗癌效果.得到了具有良好磁响应、释药温度T=41℃、可近红外成像的载药脂质体.     

Biodegradable Bi2O2Se Quantum Dots for Photoacoustic Imaging‐Guided Cancer Photothermal Therapy

As new 2D layered nanomaterials, Bi₂O₂Se nanoplates have unique semiconducting properties that can benefit biomedical applications. Herein, a facile top‐down approach for the synthesis of Bi₂O₂Se quantum dots (QDs) in a solution is described. The Bi₂O₂Se QDs with a size of 3.8 nm and thickness of 1.9 nm exhibit a high photothermal conversion coefficient of 35.7% and good photothermal stability. In vitro and in vivo assessments demonstrate that the Bi₂O₂Se QDs possess excellent photoacoustic (PA) performance and photothermal therapy (PTT) efficiency. After systemic administration, the Bi₂O₂Se QDs accumulate passively in tumors enabling efficient PA imaging of the entire tumors to facilitate imaging‐guided PTT without obvious toxicity. Furthermore, the Bi₂O₂Se QDs which exhibit degradability in aqueous media not only have sufficient stability during in vivo circulation to perform the designed therapeutic functions, but also can be discharged harmlessly from the body afterward. The results reveal the great potential of Bi₂O₂Se QDs as a biodegradable multifunctional agent in medical applications.     

Tunable Fabrication of Molybdenum Disulfide Quantum Dots for Intracellular MicroRNA Detection and Multiphoton Bioimaging

Molybdenum disulfide (MoS₂) quantum dots (QDs) (size <10 nm) possess attractive new properties due to the quantum confinement and edge effects as graphene QDs. However, the synthesis and application of MoS₂ QDs has not been investigated in great detail. Here, a facile and efficient approach for synthesis of controllable‐size MoS₂ QDs with excellent photoluminescence (PL) by using a sulfuric acid‐assisted ultrasonic route is developed for this investigation. Various MoS₂ structures including monolayer MoS₂ flake, nanoporous MoS₂, and MoS₂ QDs can be yielded by simply controlling the ultrasonic durations. Comprehensive microscopic and spectroscopic tools demonstrate that the MoS₂ QDs have uniform lateral size and possess excellent excitation‐independent blue PL. The as‐generated MoS₂ QDs show high quantum yield of 9.65%, long fluorescence lifetime of 4.66 ns, and good fluorescent stability over broad pH values from 4 to 10. Given the good intrinsic optical properties and large surface area combined with excellent physiological stability and biocompatibility, a MoS₂ QDs‐based intracellular microRNA imaging analysis system is successfully constructed. Importantly, the MoS₂ QDs show good performance as multiphoton bioimaging labeling. The proposed synthesis strategy paves a new way for facile and efficient preparing MoS₂ QDs with tunable‐size for biomedical imaging and optoelectronic devices application.     

Cathodic stripping synthesis and cytotoxity studies of glutathione-capped CdTe quantum dots

A cathodic stripping of Te precursor in the presence of Cd2+ and biocompatible glutathione (GSH) was reported for facile synthesis of lowly cytotoxic and highly luminescent CdTe quantum dots (QDs) in aqueous solution. The photoluminescence, electrogenerated chemiluminescence (ECL), toxicity, and cyto-osmosis of the QDs were evaluated to reveal their potential bio-applications. The morphology and composition of as-prepared QDs were investigated by HRTEM and powder XRD spectroscopy, which indicated that the QDs consisted of a CdTe core coated with a CdS shell. The obtained CdTe/CdS core/shell QDs possessed good crystallinity, narrow monodispersity and long-term stability. These QDs showed high fluorescence quantum yields of 49% to 63% over a broad spectral range of 540-650 nm. Efficient and stable ECL of QDs was observed on the anodic potential region upon the electrode potential cycled between 1.5 and -2.0 V versus Ag/AgCl. Furthermore, human liver cancer HepG2 cells were chosen as model cells for toxicity assay of QDs. Effects of the concentration, size, and incubation time of CdTe QDs capped with GSH or mercaptoacetic acid (MAA) on the cell metabolic viability and cyto-osmosis were evaluated. GSH-capped CdTe QDs could infiltrate cytomembrane and karyothecas, and were less cytotoxic than MAA-capped ones under the same experimental conditions. The reported CdTe QDs could be good candidates of fluorescent and ECL probes for biosensing and cell imaging.     

Biocompatible Semiconductor Quantum Dots as Cancer Imaging Agents

Approximately 1.7 million new cases of cancer will be diagnosed this year in the United States leading to 600 000 deaths. Patient survival rates are highly correlated with the stage of cancer diagnosis, with localized and regional remission rates that are much higher than for metastatic cancer. The current standard of care for many solid tumors includes imaging and biopsy with histological assessment. In many cases, after tomographical imaging modalities have identified abnormal morphology consistent with cancer, surgery is performed to remove the primary tumor and evaluate the surrounding lymph nodes. Accurate identification of tumor margins and staging are critical for selecting optimal treatments to minimize recurrence. Visible, fluorescent, and radiolabeled small molecules have been used as contrast agents to improve detection during real‐time intraoperative imaging. Unfortunately, current dyes lack the tissue specificity, stability, and signal penetration needed for optimal performance. Quantum dots (QDs) represent an exciting class of fluorescent probes for optical imaging with tunable optical properties, high stability, and the ability to target tumors or lymph nodes based on surface functionalization. Here, state‐of‐the‐art biocompatible QDs are compared with current Food and Drug Administration approved fluorophores used in cancer imaging and a perspective on the pathway to clinical translation is provided.     

Super‐Resolution Nonlinear Photothermal Microscopy

Super‐resolution fluorescence microscopy enables imaging of fluorescent structures beyond the diffraction limit. However, this technique cannot be applied to weakly fluorescent cellular components or labels. As an alternative, photothermal microscopy based on nonradiative transformation of absorbed energy into heat has demonstrated imaging of nonfluorescent structures including single molecules and ~1‐nm gold nanoparticles. However, previously photothermal imaging has been performed with a diffraction‐limited resolution only. Herein, super‐resolution, far‐field photothermal microscopy based on nonlinear signal dependence on the laser energy is introduced. Among various nonlinear phenomena, including absorption saturation, multiphoton absorption, and signal temperature dependence, signal amplification by laser‐induced nanobubbles around overheated nano‐objects is explored. A Gaussian laser beam profile is used to demonstrate the image spatial sharpening for calibrated 260‐nm metal strips, resolving of a plasmonic nanoassembly, visualization of 10‐nm gold nanoparticles in graphene, and hemoglobin nanoclusters in live erythrocytes with resolution down to 50 nm. These nonlinear phenomena can be used for 3D imaging with improved lateral and axial resolution in most photothermal methods, including photoacoustic microscopy.     

Preparation and characterization of overcoated II-VI quantum dots

A convenient route for the synthesis of high-quality overcoated II-VI quantum dots (QDs) is reported in this paper. Simple salts, such as Cd(Ac)2 and Zn(Ac)2 were used to replace organometallics, whose disadvantage is obvious. Size-tunable core/shell structured QDs (CdSe/ZnS, CdSe/CdS, etc.) were synthesized. They were of narrow size distribution and had good monodispersivity and photoluminescence (PL) properties. The spectrum was symmetrical and sharp-pointed (with the full width at half-maximum (fwhm) of about 20-30 nm). The quantum yield (QY) was improved to 60-80% from 20-30% for bare QDs and remained stable at least for 6 months. The primary overcoated QDs were modified with biomacromolecules by a direct mechanical rubbing strategy, which is very simple and fast. The results obtained by UV-vis, PL, atomic force microscopy (AFM), and fluorescence microscopy imaging showed that the modified QDs were of good fluorescent and monodisperse characteristics. They are likely to be used further for biological labels.     

Biosynthesis of biocompatible cadmium telluride quantum dots using yeast cells

We demonstrate a simple and efficient biosynthesis method to prepare easily harvested biocompatible cadmium telluride (CdTe) quantum dots (QDs) with tunable fluorescence emission using yeast cells. Ultraviolet-visible (UV-vis) spectroscopy, photoluminescence (PL) spectroscopy, X-ray diffraction (XRD), and transmission electron microscopy (TEM) confirm that the CdTe QDs are formed via an extracellular growth and subsequent endocytosis pathway and have size-tunable optical properties with fluorescence emission from 490 to 560 nm and a cubic zinc blende structure with good crystallinity. In particular, the CdTe QDs with uniform size (2-3.6 nm) are protein-capped, which makes them highly soluble in water, and in situ bio-imaging in yeast cells indicates that the biosynthesized QDs have good biocompatibility. This work provides an economic and environmentally friendly approach to synthesize highly fluorescent biocompatible CdTe QDs for bio-imaging and bio-labeling applications.      

Fluorescent nanocrystals as colloidal probes in complex fluids measured by fluorescence correlation spectroscopy

The diffusion properties of fluorescent colloidal CdSe and CdSe/ZnS nanocrystals (QDs) with different hydrophilic coatings were characterized in complex fluids such as actin solutions using fluorescence correlation spectroscopy (FCS). The hydrodynamic radii of the QDs were determined both in organic solvents and water. Attention was given to the potential artifacts arising from the fluorescence properties of the QDs. With increasing excitation intensities, the apparent particle concentration and diffusion times are overestimated if using a simple diffusion model. This can be explained by a numerical simulation. The diffusion behavior of QDs in actin networks of different concentrations was determined to demonstrate the potential use of nanocrystals as probes in soft biological matter. The decreasing diffusion coefficient of the nanocrystals with increasing actin concentration results in an intrinsic polymer viscosity of 0.12+/-0.02 ml mg(-1), in accordance with literature values.     

Aqueous‐Phase Synthesis of Highly Luminescent CdTe/ZnTe Core/Shell Quantum Dots Optimized for Targeted Bioimaging

Semiconductor quantum dots (QDs) have traditionally been synthesized in organic phase and transferred to aqueous solution by functionalizing their surface with silica, polymers, short‐chain thiol ligand, or phospholipid micelles. However, these complex steps result in i) a reduction of the quantum yield (QY) of QDs, ii) partial degrdation of the QDs, and iii) a drastic increase in the hydrodynamic size of QDs, which may hinder their biomedical applications. In this work, the fabrication and applications of cysteine‐capped CdTe/ZnTe QDs, which are directly synthesized in aqueous media, as optical probes for specific targeting of pancreatic and esophageal cancer cells in vitro are reported, as well as their capability for in vivo imaging. The CdTe/ZnTe QDs are synthesized in a one‐pot method and capped with amino acid cysteine, which contains both carboxyl and amine functional groups on their surfaces for bioconjugation. The fabricated QDs have an ultrasmall hydrodynamic diameter (3–5 nm), possess high QY (52%), and are non‐toxic to cells at experimental dosages. Confocal imaging is used to demonstrate a receptor‐mediated uptake of antibody‐conjugated QDs into pancreatic cancer cells in vitro. In vitro cytotoxicity studies (MTS‐assay) show that the IC₅₀ value of these QDs is ≈160 µg mL^?1, demonstrating low toxicity. In addition, the QDs are used for small‐animal imaging where the in vivo biocompatiblity of these QDs and their clearance following systemic injection is studied.