CdS量子点的酿酒酵母仿生合成及光谱表征

以酿酒酵母为载体,常温下利用仿生法成功合成了CdS量子点。荧光发射光谱、紫外吸收光谱以及荧光显微镜照片证明,该方法合成的CdS量子点的荧光发射峰位置在443nm,在紫外灯下能发蓝绿色荧光。透射电子显微镜(TEM)表征结果表明,该仿生法合成的CdS量子点为六方纤锌矿结构。以荧光发射和紫外吸收光谱为性能指标,考察了酿酒酵母生长时期、Cd2+的反应浓度以及反应时间等条件对合成CdS量子点的影响。当酿酒酵母处于生长稳定期初期时,与浓度为0.5mmol.L-1的Cd2+共培养24h后所合成的CdS量子点荧光最强。实验中观察到,换液培养可有效提高酿酒酵母合成CdS量子点的产量。     

Enhanced Oxidation Stability of Quasi Core-Shell Alloyed CdSeS Quantum Dots Prepared Through Aqueous Microwave Synthesis Technique

Quasi core shell alloyed CdSeS quantum dots (QDs) have been prepared through a facile aqueous-phase route employing microwave irradiation technique. The optical spectroscopy and structure characterization evidenced the quasi core shell alloyed structures of CdSeS QDs. The X-ray diffraction patterns of the obtained CdSeS QDs displayed peak positions very close to those of bulk cubic CdS crystal structures and the result of X-ray photoelectron spectroscopy data re-confirmed the thick CdS shell on the CdSe core. The TEM images and HRTEM images of the CdSeS QDs ascertained the well-defined spherical particles and a relatively narrow size distribution. On the basis, the stability of the obtained QDs in an oxidative environment was also discussed using etching reaction by H₂O₂. The experiments result showed the as-prepared QDs present high tolerance towards H₂O₂, obviously superior to the commonly used CdTe QDs and core-shell CdTe/CdS QDs, which was attributed to the unique quasi core-shell CdSeS crystal structure and the small lattice mismatch between CdSe and CdS semiconductor materials. This assay provided insight to obtain high stable crystal structured semiconductor nanocrystals in the design and synthesis process.     

Non-radiative exciton recombination through excitation energy transfer in quantum dot clusters

Excitation energy transfer (EET) processes in CdSe/CdZnS quantum dot (QD) clusters have been investigated in this study by measuring their time-resolved and spectrally resolved fluorescence intensities. The contributions of radiative and non-radiative exciton recombination through EET are evaluated, where the latter is expected to occur in a large class of QD ensembles because of the presence of nonluminescent QDs. It appears that the fluorescence decay in larger QDs serving as acceptor does not show an initial rise, in addition the lifetime of the acceptor QD is independent of the excitation wavelength, suggesting that an EET is followed mostly by non-radiative recombination.     

基于InP@ZnS QDs/Dured纳米荧光探针的DNA检测

利用巯基丙酸包覆的In P@Zn S量子点(QDs)与Dured构建了一种检测DNA的荧光探针。在该探针中,以环境友好型带负电的In P@Zn S量子点为荧光团,与带正电的Dured通过静电结合,构建了In P@Zn S QDs/Dured纳米荧光探针。通过荧光共振能量转移(FRET)机理,量子点荧光被猝灭;当DNA存在时,Dured与DNA的特异性结合使Dured从In P@Zn S QDs表面脱附,FRET过程被打断,In P@Zn S QDs荧光恢复,以荧光"关-开"方式检测DNA。该探针检测DNA的线性范围为2.0~275.0 ng·L-1,检测限为1.0 ng·L-1,并可用于模拟生物生理条件下的DNA检测。     

Cytotoxicity and fluorescence studies of silica-coated CdSe quantum dots for bioimaging applications

The toxicological effects of silica-coated CdSe quantum dots (QDs) were investigated systematically on human cervical cancer cell line. Trioctylphosphine oxide capped CdSe QDs were synthesized and rendered water soluble by overcoating with silica, using aminopropyl silane as silica precursor. The cytotoxicity studies were conducted by exposing cells to freshly synthesized QDs as a function of time (0–72 h) and concentration up to micromolar level by Lactate dehydrogenase assay, MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assay, Neutral red cell viability assay, Trypan blue dye exclusion method and morphological examination of cells using phase contrast microscope. The in vitro analysis results showed that the silica-coated CdSe QDs were nontoxic even at higher loadings. Subsequently the in vivo fluorescence was also demonstrated by intravenous administration of the QDs in Swiss albino mice. The fluorescence images in the cryosections of tissues depicted strong luminescence property of silica-coated QDs under biological conditions. These results confirmed the role of these luminescent materials in biological labeling and imaging applications.     

Prism-based Spectral Imaging of Four Species of Single-molecule Fluorophores by Using One Excitation Laser

A prism-based imaging system for simultaneously detecting four species of single-molecule (SM) fluorophores was developed. As for the detection method, four spectrally distinct species of BigDye fluorophores were bound to 50-nm-diameter gold nanoparticles (AuNPs) to form AuNP/BigDye complexes. Four species of complexes were randomly immobilized on different fused-silica slides. BigDyes were excited by an argon-ion-laser (excitation wavelengths: 488 and 514.5 nm) beam through total internal reflection on the slide surface. SM fluorescence emitted from a complex was spectrally dispersed through a prism to form an SM spot elongated in the spectral direction on a charge-coupled device. A scattered light spot generated by the AuNP of the same complex under 594-nm laser illumination was used as a wavelength reference, and the SM fluorescence spectrum was obtained from the pixel-intensity pattern of the elongated SM spot. Peak locations of fluorescence spectra of all the observed SM spots were obtained, and their histograms were distinctly separated according to species. SM spots can thus be classified as one of four species according to their peak locations. By statistically analyzing the histograms, the classification accuracy was estimated to be above 93.8 %. The number of pixels in the spectral direction required for classifying four species of SM fluorophores was estimated to be 10. As for the conventional system (which uses two excitation lasers), 15 pixels are required. Using BigDyes as the four fluorophores (which consist of donors linked to acceptors and can be excited by just an argon-ion laser) is the reason that such a small number of pixels was achieved. The developed system can thus detect 1.5 times more SM fluorophores per field of view; that is, its throughput is 1.5 times higher. The approach taken in this study, namely, using BigDye with a prism-type system, is effective for increasing the throughput of DNA microarray-chip analysis and SM real-time DNA sequencing.     

Self-assembly of dandelion-like Fe3O4@C@BiOCl magnetic nanocomposites with excellent solar-driven photocatalytic properties

In comparison with conventional organic dyes, quantum dots (QDs) have unique optical and electronic properties, which provide QDs with a wide scope of prospective application in biology and biomedicine. However, the toxicity of QDs and the fluorescence intensity of labeled bacteria must precede their application in bacterial imaging and tracing in vivo. Here, we show that treatment with CaCl₂ significantly improved bacterial labeling efficiency of CdSe/ZnS QDs with the CdSe core size of ~3.1 nm (relative fluorescence unit (RFU) value and ratio of fluorescent E. coli) with rising CdSe/ZnS QDs concentration in a concentration-dependent manner. At 12.5 nmol/L CdSe/ZnS QDs concentration, labeled Escherichia coli (E. coli) DH5α appeared as short rod-shaped and luminescent with normal size, and the survival rate and ultrastructure did not change in comparison to the control. But the ratio of fluorescent bacteria and RFU were very low. However, the survival rate of transformed E. coli was significantly inhibited by high CdSe/ZnS QDs concentrations (≥25 nmol/L). Moreover, internalization of CdSe/ZnS QDs resulted in ultrastructure damage of transformed E. coli in a concentration-dependent manner (≥25 nmol/L). Therefore, CdSe/ZnS QDs may not suitable for tracing of bacteria in vivo. Moreover, our study also revealed that colony-forming capability assay and transmission electron microscopy could be used to comprehensively evaluate the toxicity of QDs on labeled bacteria. Our findings do provide a new direction toward the improvement and modification of QDs for use in imaging and tracing studies in vivo.     

A Simple QD–FRET Bioprobe for Sensitive and Specific Detection of Hepatitis B Virus DNA

We report here a simple quantum dot-FRET (QD-FRET) bioprobe based on fluorescence resonance energy transfer (FRET) for the sensitive and specific detection of hepatitis B virus DNA (HBV DNA). The proposed one-pot HBV DNA detection method is very simple, rapid and convenient due to the elimination of the washing and separation steps. In this study, the water-soluble CdSe/ZnS QDs were prepared by replacing the trioctylphosphine oxide on the surface of QDs with mercaptoacetic acid (MAA). Subsequently, DNA was attached to QDs surface to form the functional QD-DNA bioconjugates by simple surface ligand exchange. After adding 6-carboxy-X-rhodamine (ROX)-modified HBV DNA (ROX-DNA) into the QD-DNA bioconjugates solution, DNA hybridization between QD-DNA bioconjugates and ROX-DNA was formed. The resulting hybridization brought the ROX fluorophore, the acceptor, and the QDs, the donor, into proximity, leading to energy transfer from QDs to ROX. When ROX-DNA was displaced by the unlabeled HBV DNA, the efficiency of FRET was dramatically decreased. Based on the changes of both fluorescence intensities of QDs and ROX, HBV DNA could be detected with high sensitivity and specificity. Under the optimized conditions, the linear range of HBV DNA determination was 2.5 – 30 nmol L^?1, with a correlation coefficient (R) of 0.9929 and a limit of detection (3σ black) of 1.5 nmol L^?1. The relative standard deviation (R.S.D.) for 12 nmol L^?1 HBV DNA was 0.9 % (n?=?5). There was no interference to non-complementary DNA. Time-resolved fluorescence spectra and fluorescence images were performed to verify the validity of this method and the results were satisfying.     

频谱编码深度成像实验研究

频谱编码显微镜是用衍射光栅和光谱分析装置来获得显微图像.样品上不同的位置被不同的波长照明,通过对反射光光谱进行解码来得到空间信息.搭建了一个基于超连续光源和自制光谱仪的频谱编码显微成像系统,其横向分辨率为1.72±0.13μm(编码线方向)和1.26±0.08μm(垂直于编码线方向),测得不同横向位置处的轴向分辨率有差异.对离体猪肝组织不同部位进行了成像(可见血管、肝窦内皮细胞和肝细胞);对鸡心组织以10μm深度间隔进行成像,测得不同深度处结构信息不一样.结果表明,采用该频谱编码成像的方法能够进行高分辨的深度成像.     

Size dependence of biexciton binding energy in single InAs/GaAs quantum dots

This paper studies the size dependence of biexciton binding energy in single quantum dots (QDs) by using atomic force microscopy and micro-photoluminescence measurements. It finds that the biexciton binding energies in the QDs show ``binding' and ``antibinding' properties which correspond to the large and small sizes of QDs, respectively. The experimental results can be well interpreted by the biexciton potential curve, calculated from the exciton molecular model and the Heitler--London method.     

Time-Resolved FRET and FLIM of Four-way DNA Junctions

Conformational transitions in a 4-way DNA junction when titrated with ionic solutions are studied using time-resolved fluorescence resonance energy transfer. Parameters characterising the transition in terms of critical ion concentration (c _1/2) and the Hill coefficient for ion binding are obtained by fitting a simple two-state model using steady-state spectra. Data obtained from a fluorescence lifetime plate reader and analysed by fitting a single exponential to donor fluorescence lifetime decays are shown to be in good agreement with the parameters obtained from steady-state measurements. Fluorescence lifetimes, however, offer advantages, particularly in being independent of fluorophore concentration, output intensity, inhomogeneity in the excitation source and output wavelength. We demonstrate preliminary FRET-FLIM images of DNA junction solutions obtained using a picosecond gated CCD which are in agreement with results from a fluorescence lifetime plate reader. The results suggest that time-resolved FRET-FLIM is sensitive to subtle structural changes and may be useful in assays based on 4-way DNA junctions.     

Electric field effect on the donor impurity states in zinc-blende symmetric InGaN/GaN coupled quantum dots

Based on the effective-mass approximation, the donor binding energy in a cylindrical zinc-blende (ZB) symmetric InGaN/GaN coupled quantum dots (QDs) is investigated variationally in the presence of an applied electric field. Numerical results show that the ground-state donor binding energy is highly dependent on the impurity positions, coupled QDs structure parameters and applied electric field. The applied electric field induces an asymmetric distribution of the donor binding energy with respect to the center of the coupled QDs. When the impurity is located at the center of the right dot, the donor binding energy has a maximum value with increasing the dot height. Moreover, the donor binding energy is the largest and insensitive to the large applied electric field (F?400 kV/cm) when the impurity is located at the center of the right dot in ZB symmetric In_0.1Ga_0.9N/GaN coupled QDs. In addition, if the impurity is located inside the right dot, the donor binding energy is insensitive to large middle barrier width (Lmb?2.5 nm) of ZB symmetric In_0.1Ga_0.9N/GaN coupled QDs.     

Preparation and purification of l-cysteine capped CdTe quantum dots and its self-recovery of degenerate fluorescence

l-cysteine capped CdTe quantum dots (QDs) were prepared in aqueous solution by a simple and efficient method, showing many advantages such as short synthesis period, the broaden range of starting pH value and the wide fluorescence emission wavelength range. A novel purification process was designed to remove excess Cd²⁺ which has potential cytotoxicity for bio-analysis. Three-dimensional fluorescence charts of pre- and post-purification showed that the purified QDs were of better luminescent performance. The prepared QDs were of cubic crystal structure with an average size of 2-6 nm, which were characterized by XRD and HRTEM. It is confirmed by IR spectra that the l-cysteine ligands were conjugated with CdTe cores via covalent bond. The degenerate fluorescence of QDs can be self-recovered in the presence of l-cysteine without other processing steps.     

Fluorescence Property of ZnO Nanoparticles and the Interaction with Bromothymol Blue

We synthesized ZnO quantum dots (QDs) simply in alcoholic solution, and investigated the interaction between ZnO QDs and bromothymol blue. The structural, morphological, size and spectral properties of ZnO QDs were studied. It was found that ZnO QDs were spherical nanoparticles in the crystal structure, and the average diameter of ZnO QDs was about 4.8 nm. The excitation and emission peaks were located at 346 nm and 520 nm, respectively, which were obtained on a common fluorophotometer. The quantum yield of ZnO QDs was obtained by using quinine sulfate as a reference reagent. In addition, the fluorescence of ZnO QDs can be quenched by bromothymol blue, and the quenching mechanism was proposed in a dynamic quenching mode.     

Spectrophotometric Study on the Binding of Two Water Soluble Schiff Base Complexes of Mn (III) with ct-DNA

In this work, binding of two water soluble Schiff base complexes: Bis sodium (5-sulfosalicylaldehyde) o- phenylendiiminato) Manganese (III) acetate (Salophen complex) and Bis sodium (5-sulfosalicylaldehyde) 1, 2 ethylendiiminato) Manganese (III) acetate (Salen complex) with calf thymus (ct) DNA were investigated by using different spectroscopic and electrometric techniques including UV-vis, Circular dichroism (CD) and fluorescence spectroscopy, viscommetry and cyclic voltammetry (CV). Both complexes have shown a hyperchromic and a small bathochromic shift in the visible region spectra. A competitive binding study showed that the enhanced emission intensity of ethidium bromide (EB) in the presence of DNA was quenched by the addition of the two Schiff base complexes indicating that they displace EB from its binding site in DNA. Moreover structural changes in the CD spectra and an increase in the CV spectra with addition of DNA were observed. The results show that both complexes bind to DNA. The binding constants have been calculated using fluorescence data for two complexes also K_b was calculated with fluorescence Scatchard plot for Salophen. Ultimately, the experimental results show that the dominant interactions are electrostatic while binding mode is surface binding then followed by hydrophobic interactions in grooves in high concentration of complexes.     

Preparation of water-soluble CdTe/CdS core/shell quantum dots with enhanced photostability

CdTe/CdS core/shell quantum dots (QDs) have been synthesized in an aqueous phase using thioacetamide as a sulfur source. The quantum yield was greatly enhanced by the epitaxial growth of a CdS shell, which was confirmed by X-ray photoelectron spectroscopy (XPS) results. The quantum yield of as-prepared CdTe/CdS core/shell QDs without any post-preparative processing reached 58%. The experimental results illustrate that the QDs with core/shell structure show better photostability than thioglycolic acid (TGA)-capped CdTe QDs. The cyclic voltammograms reveal higher oxidation potentials for CdTe/CdS core/shell QDs than for TGA-capped CdTe QDs, which explains the superior photostability of QDs with a core/shell structure. This enhanced photostability makes these QDs with core/shell structure more suitable for bio-labeling and imaging.     

Analysis of Cation-Dependent DNA (G<Subscript>3</Subscript>T<Subscript>1</Subscript>)<Subscript>4</Subscript> Shape Change Using Fluorescence Correlation Spectroscopy

In G-rich DNA, it is well known that the form changes from single-strand DNA to G-quadruplex due to cations. In this study, we analyze the diffusion coefficient and fluorescence intensity obtained by fluorescence correlation spectroscopy for short G-rich DNA of the (G₃T₁)₄ sequence labeled as 5-Carboxytetramethylrhodamine (TAMRA) with variation of the K⁺ ion concentration. At a K⁺ ion concentration of more than 200 mM, the single-strand DNA was changed to the G-quadruplex. The size of the G-quadruplex decreased to 86% than the size of the single strand DNA at K⁺ ion concentration of 0 M. The size of the G-quadruplex and the fluorescence intensity of TAMRA attached to the DNA were constant with an increase in the K⁺ ion concentration between 200 and 800 mM. This means that the size of the DNA and the fluorescence intensity of the TAMRA are not affected by the K⁺ ion concentration at the G-quadruplex structure because the binding structure of DNA and TAMRA dye leads to stability at a concentration of less than 100 mM K⁺. Based on our short G-rich DNA results, longer G-rich DNA is analyzed for the diffusion coefficient of the DNA and the fluorescence intensity variation of fluorescence dye attached to the DNA.     

拉直的单个DNA分子的全内反射荧光实时成像研究

全内反射荧光(TIRF)成像技术利用穿透深度仅200 nm左右的隐失波来激发诱导荧光,探测灵敏度和图像信噪比大大提高,成为单分子研究的有力工具。分子梳技术利用DNA末端与固体表面的结合力和周围流体流动产生的侧向力将DNA分子拉伸并平铺在表面上。结合这两种技术,对分子梳拉直的单个DNA分子进行了清晰的实时荧光成像,发现TIRF成像条件下DNA分子与荧光探针YOYO-1组成的复合体可自然避免发生光敏断裂现象;实时监测了单个DNA-YOYO-1复合体的光漂白过程,通过对激发光照射时间与探测器曝光时间进行同步控制,可大幅降低光漂白程度,为拉直的单个DNA分子的长时间实时观察和成像研究优化了实验条件,为实时、可视化地研究其与蛋白质相互作用的动力学过程奠定了基础。