Functionalized quantum dots to quantify NADPH and their use for NADP+-dependent biocatalyzed transformations

Quantum dots are semiconducting nanoparticles that can be prepared with interesting optical properties. The fluorescent properties of quantum dots are one of the key advantages for their use as optical labels for biorecognition events and biocatalytic processes. We have prepared semiconductor quantum dots conjugated with Nile Blue (NB), and demonstrate that NB-functionalized quantum dots can act as versatile probes to analyze different biocatalyzed transformations, and can be used for the quantitative detection of NADPH as well as NADH. This approach provides a new path for the optical detection of NAD(P)H and for the quantitative analysis of NAD(P)(+)-dependent biotransformations.     

Noninvasive imaging of quantum dots in mice

Quantum dots having four different surface coatings were tested for use in in vivo imaging. Localization was successfully monitored by fluorescence imaging of living animals, by necropsy, by frozen tissue sections for optical microscopy, and by electron microscopy, on scales ranging from centimeters to nanometers, using only quantum dots for detection. Circulating half-lives were found to be less than 12 min for amphiphilic poly(acrylic acid), short-chain (750 Da) methoxy-PEG or long-chain (3400 Da) carboxy-PEG quantum dots, but approximately 70 min for long-chain (5000 Da) methoxy-PEG quantum dots. Surface coatings also determined the in vivo localization of the quantum dots. Long-term experiments demonstrated that these quantum dots remain fluorescent after at least four months in vivo.     

Plasmonic Control of Spontaneous Emission of Quantum Dots in Sub-Wavelength Photonic Templates

Finite difference time domain (FDTD) simulations were performed on two different plasmonic sub-wavelength photonic templates embedded with CdSe quantum dots. Tunable loading of these templates with plasmonic nano antenna allowed control of the emission from the embedded quantum dots. We discuss how large loading of nano antenna can effectively control the optical density of states for the quantum dots leading to enhancement of their radiative decay rates as observed in experiments. On the other hand, at low level of loading, while FDTD fails to capture the observed enhancement of decay rates in experiment, an alternative mechanism is suggested to exist in such cases. Thus, subtle interplay of multiple mechanisms engineered by appropriate placement and loading of plasmonic nano antenna in such templates is demonstrated as an effective method to control optical density of states and hence spontaneous emission of embedded quantum dots.     

量子点在生物学中的研究进展

量子点作为一种新型的荧光标记物近年来已在生物学中获得广泛应用。本文总结了量子点的主要光学特性,其中包括荧光激发和发射光谱特性、量子产额、光漂白特性和荧光寿命等。重点综述了量子点在细胞标记、活体和组织成像、组合标记和光动力学治疗等生物学中的应用及其最新研究进展。同时讨论了量子点在应用中可能存在的细胞毒性等主要问题,最后对量子点在生物学中的应用前景作了展望。     

Luminescent quantum dots for multiplexed biological detection and imaging

Recent advances in nanomaterials have produced a new class of fluorescent labels by conjugating semiconductor quantum dots with biorecognition molecules. These nanometer-sized conjugates are water-soluble and biocompatible, and provide important advantages over organic dyes and lanthanide probes. In particular, the emission wavelength of quantum-dot nanocrystals can be continuously tuned by changing the particle size, and a single light source can be used for simultaneous excitation of all different-sized dots. High-quality dots are also highly stable against photobleaching and have narrow, symmetric emission spectra. These novel optical properties render quantum dots ideal fluorophores for ultrasensitive, multicolor, and multiplexing applications in molecular biotechnology and bioengineering.     

Nondestructive quantum dot-based intracellular serotonin imaging in intact cells

In recent years, quantum dots (Qdot), with their unique physical, chemical, and optical properties, have been used extensively as probes to visualize several cell membrane receptors and extracellular biomolecules. However, Qdot-based intracellular imaging has always been associated with vital lacunas. High affinity between quantum dots may induce serious aggregation in the cytoplasm; as a result, quantum dot aggregates are usually misinterpreted as quantum dot-probed intracellular molecules. Moreover, due to the more viscous nature of the cytoplasm versus the extracellular aqueous media, aggregation issues become more severe during intracellular studies. In this work, we suggest direct nondestructive serotonin imaging in an intact cell using the quantum dot-based immunoassay with a rapid tunable multicolor imaging system based on the acousto-optic tunable filter. Any false-positive intracellular serotonin molecules that appeared due to the aggregation of quantum dots could be completely discriminated from the real intracellular serotonin granules through multicolor cellular imaging. The developed method is quick and has wide applicability in targeting various intracellular proteins, coenzymes, and micronutrients.     

含镉量子点的毒性研究进展

含镉量子点是典型的量子点,近年来受到广泛研究。含镉量子点的潜在毒性是其在生物成像及生物医药方面应用和发展的关键制约因素,因此,对其毒性作用的研究具有重要意义。目前对含镉量子点的体外毒性研究主要集中在人肝癌细胞(HepG2)、神经分泌细胞(PC12)等细胞实验及斑马鱼胚胎体外培养实验。体内毒性研究包括小鼠等动物实验。这些研究证实,量子点对HepG2等细胞系和小鼠、贻贝等动物均具细胞毒性。研究者们普遍认为,量子点是通过释放其组成中的重金属,诱导生物体产生活性氧自由基,进而引发细胞凋亡或自噬,但对量子点的具体毒性作用机制并不完全清楚。该文对含镉量子点的体内和体外毒性研究工作进展进行了综述,包括含镉量子点对肝肾细胞、神经细胞、血液细胞及免疫细胞等体外毒性研究工作,对陆生及水生动物等的体内毒性研究工作,旨在更好、更全面地评估含镉量子点的毒性,为今后对量子点的毒性作用机制研究提供方向,促进含镉量子点在生物医学方面的发展和应用。     

量子点的制备及其在生物医学中的研究

量子点是近几年发展起来的新型纳米材料,虽然研究起步较晚,但因其独特的电学和光学性质而成为人们关注的热点,在生物医学等多个领域有突破性的研究进展。本文主要介绍量子点的性质、制备方法及其在生物医学中的应用进展和存在的问题。     

Water-soluble quantum dots for biomedical applications

Semiconductor nanocrystals are 1-10nm inorganic particles with unique size-dependent optical and electrical properties due to quantum confinement (so they are also called quantum dots). Quantum dots are new types of fluorescent materials for biological labeling with high quantum efficiency, long-term photostability, narrow emission, and continuous absorption spectra. Here, we discuss the recent development in making water-soluble quantum dots and related cytotoxicity for biomedical applications.     

体内光量子点可视化图像在脑肿瘤中的应用

恶性胶质瘤年发病率约为5/100,000。美国每年有超过14,000例的新发恶性脑胶质瘤患者。治疗主要以手术治疗为主,手术肿瘤的切除程度影响患者的预后。外科手术治疗脑肿瘤需要精确定位脑肿瘤组织在正常脑组织中的位置以便能够获得精确的组织活检和肿瘤的完全切除。量子点是稳定存在的,产生荧光的可视化半导体纳米晶体。静脉注射量子点伴随着网状内皮系统和巨噬细胞的隔离。巨噬细胞可渗入到肿瘤组织并且能够吞噬通过静脉注射的光量子来产生可视化的肿瘤标记。通过巨噬细胞介导,将光量子运输至肿瘤组织展现了一种新兴技术来标记术前肿瘤组织。由于肿瘤组织中的光量子可以被光学成像和光谱学工具来探测,因此在脑肿瘤组织活检和切除中可以为外科医生提供可视化得实时反馈。     

CdTe quantum dots and YAG hybrid phosphors for white light‐emitting diodes

CdTe quantum dots, 3.28 nm in size, were synthesized using a one‐step method in an aqueous medium. The CdTe quantum dots were successfully employed as hybrid phosphors for white light‐emitting diode (LED) devices by combining them with yellow‐emitting YAG:Ce phosphor. The color‐rendering index value and International Commission on illumination coordinates for hybrid phosphor white LEDs were 75 and (x = 0.30, y = 0.29), respectively. Compared with conventional phosphors, semiconductor quantum dots have larger band gap energy and broader absorption features, and can be excited more efficiently by optical pumping sources. The results confirmed that the high color‐rendering index value of the white LED was due to the CdTe quantum dots introduced in the hybrid phosphor system. Copyright © 2013 John Wiley & Sons, Ltd.     

Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications

Fluorescent nanocrystals, specifically quantum dots, have been a useful tool for many biomedical applications. For successful use in biological systems, quantum dots should be highly fluorescent and small/monodisperse in size. While commonly used cadmium-based quantum dots possess these qualities, they are potentially toxic due to the possible release of Cd²⁺ ions through nanoparticle degradation. Indium-based quantum dots, specifically InP/ZnS, have recently been explored as a viable alternative to cadmium-based quantum dots due to their relatively similar fluorescence characteristics and size. The synthesis presented here uses standard hot-injection techniques for effective nanoparticle growth; however, nanoparticle properties such as size, emission wavelength, and emission intensity can drastically change due to small changes in the reaction conditions. Therefore, reaction conditions such temperature, reaction duration, and precursor concentration should be maintained precisely to yield reproducible products. Because quantum dots are not inherently soluble in aqueous solutions, they must also undergo surface modification to impart solubility in water. In this protocol, an amphiphilic polymer is used to interact with both hydrophobic ligands on the quantum dot surface and bulk solvent water molecules. Here, a detailed protocol is provided for the synthesis of highly fluorescent InP/ZnS quantum dots that are suitable for use in biomedical applications.     

Creating self-illuminating quantum dot conjugates

Semiconductor quantum dots are inorganic fluorescent nanocrystals that, because of their unique optical properties compared with those of organic fluorophores, have become popular as fluorescent imaging probes. Although external light excitation is typically required for imaging with quantum dots, a new type of quantum dot conjugate has been reported that can luminesce with no need for external excitation. These self-illuminating quantum dot conjugates can be prepared by coupling of commercially available carboxylate-presenting quantum dots to the light-emitting protein Renilla luciferase. When the conjugates are exposed to the luciferase's substrate coelenterazine, the energy released by substrate catabolism is transferred to the quantum dots through bioluminescence resonance energy transfer, leading to quantum dot light emission. This protocol describes step-by-step procedures for the preparation and characterization of these self-illuminating quantum dot conjugates. The preparation process is relatively simple and can be done in less than 2 hours. The availability of self-illuminating quantum dot conjugates will provide many new possibilities for in vivo imaging and detection, such as monitoring of in vivo cell trafficking, multiplex bioluminescence imaging and new quantum dot-based biosensors.     

量子点在新药开发中的应用

由于具有优异的光学特性,量子点在生物医学领域内的研究和应用取得了一些有意义的进展,同时也引起了新药开发人员的兴趣.本文概述了量子点在新药开发中所具有的优势,分析了量子点在药物传输、药物筛选和药靶确证方面的潜在应用,进一步讨论了当前量子点应用于新药开发存在的问题和不足.     

From analog to digital: exploring cell dynamics with single quantum dots

Semiconductor quantum dots (QDs) have emerged as new fluorescent probes for biology. When combined with ultrasensitive optical techniques, they allow motions of individual biomolecules to be tracked in live cells with high signal-to-noise and over unprecedented durations. Single QD imaging readily offers a powerful tool to investigate the organization in cell membranes. Altogether QDs will contribute to more advanced biological imaging and enable new studies on the dynamics of cellular processes.Robert Feulgen Lecture 2005 presented at the Joint Meeting of the Society for Histochemistry and The Histochemical Society in Noordwijkerhout, The Netherlands     

Quantification of tyramine in different types of food using novel green synthesis of ficus carica quantum dots as fluorescent probe

Tyramine (TYM) is catecholamine releasing compound and TYM rich food causes hypertensive crisis due to a combination with monoamine oxidase inhibitor (MAOIs). Therefore, analysis of TYM in TYM rich food as (old cheese, cured meat, sausage, pickled olive and canned fish) and the environment is essential for hypertensive patients and to improvement of food industries. In this work, TYM was analyzed in different types of food using novel green synthesis carbon dots from Ficus carica (fig fruits). The gradual addition of TYM to polyamine carbon quantum dots (PA@CQDs) led to enhancement of the quantum dots fluorescence due to formation of hydrogen bonding between quantum dots and TYM. The calibration graph was plotted in the range 5–400 ng mL⁻¹. The method was applied for the determination of TYM in different types of food as old cheese, cured meat, sausage, pickled olive and canned fish. The lower limit of quantitation (LOQ) was found to be 1.68 ng mL⁻¹. The method was successfully applied for the quantification of TYM in varying types of food with high sensitivity and high economic effect due to the reusability of the quantum dots. The optical and morphological characters of quantum dots were studied carefully.     

Quantum dots-bienzyme hybrid system for the sensitive determination of glucose

In this paper, we attempt to develop a sensitive detection method for glucose with the combination of the unique optical property of quantum dots and the specificity of enzymatic reactions. With glucose and hydroquinone as substrates, benzoquinone that intensively quenches the photoluminescence of quantum dots can be produced via the catalysis of bienzyme (glucose oxidase and horseradish peroxidase) system. A relatively low detection limit of 1.0x10(-8)mol/L can be achieved. Two linear ranges from 1.0x10(-6) to 1.5x10(-4)M and from 1.5x10(-4) to 1.0x10(-3)M were obtained. For the detection of 1.0x10(-4)M glucose, six replicative measurements showed the reproducibility (R.S.D.) of 4.43%. The quantum dots-enzymes system possesses advantages of simple procedure without modification of quantum dots or immobilization of enzymes, low cost, high sensitivity and short detection times within several minutes.     

Mortalin imaging in normal and cancer cells with quantum dot immuno-conjugates

Quantum dots are the nanoparticles that are recently emerging as an alternative to organic fluorescence probes in cell biology and biomedicine, and have several predictive advantages. These include their ⑴broad absorption spectra allowing visualization with single light source, ⑵exceptional photo-stability allowing long term studies and ⑶narrow and symmetrical emission spectrum that is controlled by their size and material composition. These unique properties allow simultaneous excitation of different size of quantum dots with a single excitation light source, their simultaneous resolution and visualization as different colors. At present there are only a few studies that have tested quantum dots in cellular imaging. We describe here the use of quantum dots in mortalin imaging of normal and cancer cells. Mortalin staining pattern with quantum dots in both normal and cancer cells mimicked those obtained with organic florescence probes and were considerably stable.     

Real-time nanoscopy by using blinking enhanced quantum dots

Superresolution optical microscopy (nanoscopy) is of current interest in many biological fields. Superresolution optical fluctuation imaging, which utilizes higher-order cumulant of fluorescence temporal fluctuations, is an excellent method for nanoscopy, as it requires neither complicated optics nor illuminations. However, it does need an impractical number of images for real-time observation. Here, we achieved real-time nanoscopy by modifying superresolution optical fluctuation imaging and enhancing the fluctuation of quantum dots. Our developed quantum dots have higher blinking than commercially available ones. The fluctuation of the blinking improved the resolution when using a variance calculation for each pixel instead of a cumulant calculation. This enabled us to obtain microscopic images with 90-nm and 80-ms spatial-temporal resolution by using a conventional fluorescence microscope without any optics or devices.     

Molecular profiling of single cancer cells and clinical tissue specimens with semiconductor quantum dots

Semiconductor quantum dots (QDs) are a new class of fluorescent labels with broad applications in biomedical imaging, disease diagnostics, and molecular and cell biology. In comparison with organic dyes and fluorescent proteins, quantum dots have unique optical and electronic properties such as size-tunable light emission, improved signal brightness, resistance against photobleaching, and simultaneous excitation of multiple fluorescence colors. Recent advances have led to multifunctional nanoparticle probes that are highly bright and stable under complex in vitro and in vivo conditions. New designs involve encapsulating luminescent QDs with amphiphilic block copolymers, and linking the polymer coating to tumor-targeting ligands and drug-delivery functionalities. These improved QDs have opened new possibilities for real-time imaging and tracking of molecular targets in living cells, for multiplexed analysis of biomolecular markers in clinical tissue specimens, and for ultrasensitive imaging of malignant tumors in living animal models. In this article, we briefly discuss recent developments in bioaffinity QD probes and their applications in molecular profiling of individual cancer cells and clinical tissue specimens.