量子点的荧光特性在生物探针方面的应用

量子点具有传统有机荧光染料无可比拟的光学魅力,在生物医学及材料领域已引起广泛的兴趣,许多科学工作者在量子点用于生物学领域方面已经取得一定进展。目前,量子点最有前途的应用领域是在生物体系中作为荧光标记物。通过观察量子点标记分子与靶分子相互作用的部位,及其在活细胞内的运行轨迹,可能为信号传递的分子机制提供线索,从而为阐明细胞生长发育的调控及癌变规律提供直观依据。文章介绍了量子点研究生物大分子之间的相互作用、生物大分子荧光标记、细胞及生物组织的荧光标记与成像以及活体成像等方面的应用。并概述了纳米量子点作为生物荧光探针的应用前景以及亟待解决的问题。     

Optimization of the Near-Infrared Fluorescence Labeling for In Vivo Monitoring of a Protein Drug Distribution in Animal Model

The objective of this study is to optimize the parameters in labeling near-infrared (NIR)fluorescent dye cypate to protein drugs for in vivo optical imaging of drug distributions in animal model. l-ASparaginase (l-ASNase) was used as a protein drug model for the study. To achieve this goal, various labeling conditions, including different catalysts, feed ratios of all components, pH conditions, temperatures, and reacting durations, were investigated. The dye-to-protein (D/P) ratio and enzymatic activity were designated as the metric to evaluate the labeling process. The stability of the cypate–protein conjugate in blood serum and its distribution in small animals were subsequently inspected. Results showed that feed ratio of l-ASNase and the pH value played the most important role in adjusting the labeling efficiency. Reaction duration and temperature had less effect on the dye-to-protein labeling properties. The optimal condition for the labeling of cypate to l-ASNase was 4 h reaction duration at 4 °C and pH 8.5 under catalysis by HOBt/HBTU. The dynamic distribution in animal model displayed that the labeled l-ASNase firstly accumulated in liver and cleared from the enteron system. This study demonstrated that the NIR image system combined with NIR probe has the capability to trace the dynamics of protein drugs in animals for drug development.     

量子点光谱成像技术与重建仿真

为满足机载星载平台对光谱成像系统紧凑型和轻量化的需求,克服当前光谱成像技术分光系统结构复杂、成本高的不足,提出了基于量子点材料的光谱成像技术方案。将条带状的量子点阵列片放置于前置镜焦面前,利用量子点材料对光谱的吸收特性对探测目标的入射光谱进行调制,使用最小二乘法建立探测目标的光谱重建模型,采用推扫的方式获取数据并进行光谱重建可以获得目标光谱和空间信息。量子点光谱成像技术具有光谱分辨率高、能量利用率高、体积小、光谱范围宽和成本低等优势。分析了不同光谱谱段间隔和噪声等因素对重建光谱分辨率的影响,以及对重建光谱准确性或者失真度的影响。分析得出谱段间隔越低,光谱分辨率越高;重建的准确性和分辨率随着噪声水平的增大而降低;适当的提高谱段间隔,可以提高重建的准确性。将已知数据立方体和它的仿真结果进行对比,可以看出还原得到的量子点光谱图像质量较好,验证了该技术的可行性。量子点材料为光谱成像技术提供了新的途径,在航空航天等小型化光谱遥感领域具有极大的应用潜力。     

Development and Investigation of Ultrastable PbS/CdS/ZnS Quantum Dots for Near‐Infrared Tumor Imaging

Achieving bright, reliable, robust, and stable probes for in vivo imaging is becoming extremely urgent for the cancer imaging research community. To date very few works have reported on elucidating in the varied and chemically complex biological milieu. The authors report detailed investigations of the synthesis of near‐infrared, water dispersive, strongly luminescent, and highly stable PbS/CdS/ZnS core/shell/shell quantum dots (QDs). These QDs are extremely stable, they could keep their initial morphology, dispersion status, and photoluminescence (PL) in phosphate buffered saline buffer for as long as 14 months. The QDs also show excellent photostability and could keep ≈80% of their initial PL intensity after 1 h continuous, strong UV illumination. More interestingly, they show negligible toxicity to cultured cells even at high QDs concentration. Given these outstanding properties, the QDs are explored for in vivo, tumor imaging in mice. With one order of magnitude lower QD concentration (0.04 mg mL^–1), significantly weaker laser intensity (0.04 W cm^–2 vs ≈1 W cm^–2), and considerably shorter signal integration time (≤1 ms vs hundreds of ms) as compared to the best reported rare earth doped nanoparticles, the QDs show high emission intensity even at injection depth of ≈2.5 mm.     

Analyzing Increased Fluorescence of Single Quantum Dots on Dehydrated Agarose by Single-Molecular Imaging

A single-molecule imaging technique was used to study emission behaviors of quantum dots (QDs) on the surface of glass and dehydrated agarose. The luminescence intensity of single QDs on the dehydrated agarose film was 2 or 3 times that on the glass. Our results revealed that the roughness and the hydrophobic nature of dehydrated agarose gel can suppress the blinking of single QDs. A clear decrease in the power law exponent for “on” time but an increase for “off” time was observed when the glass was replaced with 4% and 8% dehydrated agarose. The enhanced luminescence of QDs on dehydrated agarose will make their applications on microarrays more attractive.     

胶体水相法制备高量子产率及高稳定性的水溶性ZnSe量子点

采用简便的胶体水相法制备了高荧光强度且稳定性良好的ZnSe量子点(ZnSe QDs),克服了以往水相合成法稳定性差、量子产率低等缺陷。优化后的最佳合成条件为：以还原型L-谷胱甘肽作为稳定剂,L-谷胱甘肽∶Se2-∶Zn2+摩尔比为5∶1∶5,介质pH 10.5,反应温度在90～100 ℃之间。且合成后不需要采取任何光照后处理,ZnSe QDs的量子产率(QYs) 即可高达50.1%,放置3个月后荧光强度基本不变,水溶性优良。用紫外-可见分光光度法(UV-Vis)、荧光分光光度法(FL)、透射电子显微镜(TEM)等分析检测手段,对得到的ZnSe QDs的性能进行表征。合成的量子点在300 nm激发下发蓝紫色荧光(370 nm),其优良的光化学特性将有利于其在光热器件的制造及化学生物领域的应用。     

Enhancement of Intracellular Delivery of CdTe Quantum Dots (QDs) to Living Cells by Tat Conjugation

Quantum dots (QDs), as novel fluorescence probes, have shown a great potential for bio-molecular labeling and cellular imaging. To stain cellular targets, the sufficient intracellular delivery of QDs is required. In this work the tat, a typical membrane-permeable carrier peptide, was conjugated with thiol-capped CdTe QDs to form CdTe Tat-QDs, and the intracellular deliveries of CdTe QDs or CdTe Tat-QDs were compared in human hepatocellular carcinoma (QGY) cells and human breast cancer (MCF7) cells in vitro by means of confocal laser scanning microscopy. Added into the cell dishes, both QDs and Tat-QDs adhered to the outer leaflet of the plasma membrane of cells within a few minutes, but the binding amount of Tat-QDs was obviously higher than that of QDs. Then both QDs and Tat-QDs can penetrate into cells, and their cellular contents increased with incubation time but both saturated after 3 hours incubation. However the cellular levels of Tat-QDs were higher than those of QDs, with the ratio of 2.1 (±0.3) times in QGY cells and 1.5 (±0.2) times in MCF7 cells, demonstrating the enhancing effect of Tat conjugation on the intracellular delivery of QDs.     

油溶性CdSe量子点荧光探针直接检测农药水胺硫磷

基于农药水胺硫磷对油溶性CdSe量子点荧光猝灭的现象,建立了一种简单、快速、直接检测农药水胺硫磷的新方法。在最佳实验条件下,油溶性CdSe量子点的荧光猝灭程度与水胺硫磷浓度在2.30×10-7～1.09×10-5 mol·L-1范围内呈较好的线性关系,相关系数为0.999 9,对水胺硫磷的检出限为1.1×10-7 mol·L-1。本法已成功用于大米和小麦面粉样品中水胺硫磷农药残留的检测,加入回收率在93.3%～105.0%之间,结果满意。结合紫外-可见吸收光谱及时间分辨荧光光谱,探讨了水胺硫磷对油溶性CdSe量子点荧光猝灭的机理。研究结果表明,水胺硫磷能有效改变油溶性CdSe量子点的表面状态,增大了表面缺陷和非辐射重组的发生,从而使油溶性CdSe量子点的荧光猝灭。     

CdTe量子点荧光猝灭法测定微量铅

以CdTe量子点作为荧光探针,基于荧光猝灭法对铅进行了定量检测,考察了缓冲溶液体系、量子点浓度、反应时间等多种因素的影响。实验结果表明,在pH 7.5的0.2mol/L Na2HPO4-NaH2PO4缓冲液中,反应时间为10min,铅浓度为2.0×10-6—3.5×10-5mol/L范围时,其线性回归方程为ΔF=26.35+11.47C(×10-6mol/L),相关系数和检出限分别为0.9991和1.8×10-8mol/L。该方法灵敏度高,为铅的测定提供了新的方法。     

CdTe量子点荧光猝灭法测定钴

以CdTe量子点作为荧光探针,基于荧光猝灭法对钴(Ⅱ)进行了定量检测,考察了缓冲溶液体系、量子点浓度、反应时间等多种因素的影响。实验结果表明,在pH8.0的0.2mol/LNa2HPO4-NaH2PO4缓冲液中,反应时间为10min,钴(Ⅱ)浓度为1.6×10-5—20×10-5mol/L范围时,其线性回归方程为F0/F=1.45+0.096Q(10-5mol/L),检出限为3×10-7mol/L。该方法检测范围宽,灵敏度高,为钴的测定提供了新的方法。     

The Influence of Surface Trapping and Dark States on the Fluorescence Emission Efficiency and Lifetime of CdSe and CdSe/ZnS Quantum Dots

To investigate the influence of surface trapping and dark states on CdSe and CdSe/ZnS quantum dots (QDs), we studied the absorption, fluorescence intensity and lifetime by using one-and two-photon excitation, respectively. Experimental results show that both one- and two-photon fluorescence emission efficiencies of the QDs enhance greatly and the lifetime increase after capping CdSe with ZnS due to the effective surface passivation. The lifetime of one-photon fluorescence of CdSe and CdSe/ZnS QDs increase with increasing emission wavelength in a supralinear way, which is attributed to the energy transfer of dark excitons. On the contrary, the lifetime of two-photon fluorescence of bare and core-shell QDs decrease with increasing emission wavelength, and this indicates that the surface trapping is the dominant decay mechanism in this case.     

A Feasible and Quantitative Encoding Method for Microbeads with Multicolor Quantum Dots

Multicolor encoded beads were achieved by incorporating two color core-shell quantum dots (QDs) (CdSe/ZnS) to commercial polystyrene (PS) beads. By controlling the concentration ratios of the two quantum dots (QDs) in doping solutions, a series of codes with different intensity ratios were obtained. Based on the multiple encoded carboxylic modified polystyrene beads, fluorescent dyes labeled antibodies were distinguished successfully on the beads’ surface. It suggests that the encoded beads from this method have the practicability in biological applications and chemical analysis. Hai-Qiao Wang and Zhen-Li Huang authors contribute equally to this work     

单质碘对水溶性CdTe量子点的荧光猝灭研究

合成了不同尺寸的、巯基丙酸和半胱氨酸修饰的水溶性CdTe量子点,并研究了I2与CdTe量子点的相互作用,实验发现I2能够明显猝灭CdTe量子点的荧光,且猝灭程度与量子点的修饰剂有关,没有表现出明显的尺寸依赖效应。     

功能性CdTe量子点荧光增敏法测定对苯二胺

以CdT e量子点作为荧光探针,基于荧光增敏法对对苯二胺进行了定量检测,考察了缓冲溶液体系、量子点浓度、反应时间等多种因素的影响。实验结果表明,在pH 7.6的0.2m ol/LN a2HPO4-N aH2PO4缓冲液中,反应时间为15m in,对苯二胺浓度为6.0×10-6—1.8×10-5m ol/L范围时,其线性回归方程为ΔF=75.64＋7.95C（10-6m ol/L）,相关系数和检出限分别为0.9989和2.1×10-8m ol/L。该方法检出限低,灵敏度高,为对苯二胺的测定提供了新的选择。     

红外热成像与近红外光谱结合快速检测潜育期番茄花叶病

现有的番茄花叶病无损检测方法无法在潜育期内,即显症之前进行早期识别导致施药不及时或者盲目过度施药。设计与试制了红外热成像信息采集系统,主要包括：光箱、红外热成像仪、温度及升降控制器、加热板和升降载物台。该系统能够根据温度起止节点的要求,人为调节拍摄温度。在江苏大学现代农业装备与技术省部共建重点实验室Venlo型温室中进行非抗病性番茄品种辽宁农科院L-402的培育。采用叶面摩擦接种花叶病毒(Tobacccco mosaic virus, ToMV),分为轻度感染组(LI),重度感染组(SI);LI组为磷酸缓冲液稀释500倍后的病毒液接种,SI组为病毒原液接种。对照组(CG)喷施等量磷酸缓冲液。接种10 d后叶片开始出现病斑,证明接种后9 d为番茄花叶病的潜育期。使用红外热成像系统采集了三个组共计144个样本的红外热成像图,计算叶表最大温差(MTD) 以表征潜育期内连续9 d内的叶面温度变化情况。CG组叶片的MTD值差异极小,而接种后叶片MTD值随着病毒侵染时间的推进发生了显著的变化。接种6 d后MTD值差异最大可达1.63 ℃,第7 d开始差异逐步缩小,表明病毒的扩散范围增大导致病叶越来越多的区域被侵染使得整体叶温上升。光谱采集采用两种方法进行,一种是根据热像图的MTD值计算判别出温度突变区域后采集光谱,记为热像采集法(TCM);另一种是不考虑病灶位置,在叶尖、叶中、叶基三个区域分别随机选择一个点采集光谱后求平均值,记为随机采集法(RCM)。TCM确定三个光谱采集点的选择原则是：LI组接种后3,6和9 d的温度突变区域平均MTD值比CG组温度分别高出0.3,0.7和0.5 ℃。SI组接种后3,6和9 d的温度突变区域平均MTD值比CG组温度分别高出0.5,1.2,0.8 ℃。差值达到此标准的病灶位置才定为TCM的可选区域。对所有样本采用支持向量机(SVM)算法建立识别模型。采用主成分分析对2 151个波长点的光谱信息进行压缩,前6个主成分所对应的累积方差贡献率已到达99%。分别对感病3,6和9 d的样本按照2∶1的比例划分校正集和预测集,对预测集样本的病害程度进行识别。两种方法所建立的模型的总识别率分别为92.59%和99.77%。采用TCM建立的光谱识别模型中仅有接种后3 d的一个LI组样本未能识别出来,被误判成CG组样本外,其余组识别率均达到了100%。结果表明近红外光谱法识别番茄花叶病是可行的。采用红外热成像结合近红外光谱法能够建立识别率更高的番茄花叶病潜育期识别模型,克服点源采样随机性,对后续管控流程和突破作物早期精准用药的关键技术探索,建立更为精准的温室智能施药系统提供了新的思路。     

多谱段短波红外小鼠静脉荧光观测成像研究

短波红外(short-wave infrared,SWIR)一般指900～1700 nm的光波段,是肉眼不可见的光波段,这种波段目前主流的探测器以InGaAs为主,主要用于军事、生物以及材料光谱分析等领域.短波红外荧光成像以其对生物组织光学损伤小、成像深度大、成像信噪比高、空间和时间成像分辨率高等特点,使得基于InGa...     

巯基乙胺稳定的CdTe量子点荧光猝灭法测定间苯二酚

以CdTe量子点作为荧光探针,基于荧光猝灭法对间苯二酚进行了定量检测,考察了缓冲溶液体系、量子点浓度、反应时间等多种因素的影响.实验结果表明,在pH 6.5的0.2mol/LNa<,2>HPO<,2>-NaH<,2>PO<,4>缓冲液中,反应时间为15min.间苯二酚浓度为1.6×10<'-6>-2.0×10<'-5>...     

近红外漫射光层析成像实验研究

实验验证了所建立的时间分辨光学层析成像测量系统的光学参量预测能力、多个测量通道的一致性,表明了本测量系统具有层析成像的潜力.研究了采用特征数据量（平均飞行时间、广义脉冲谱技术为基础的用数据类型R）及全时间分辨数据的图像重建算法在光学参量重建准确度、重建出的对象尺寸、空间分辨率等方面的各自优势.     

Near-Infrared Imaging Using a High-Speed Monitoring Near Infrared Hyperspectral Camera(Compovision)

This review paper reports near-infrared (NIR) imaging studies using a newly-developed NIR camera, Compovision. Compovision can measure a significantly wide area of 150 mm×250 mm at high speed of between 2 and 5 s. It enables a wide spectral region measurement in the 1 000～2 350 nm range at 6 nm intervals. We investigated the potential of Compovision in the applications to industrial problems such as the evaluation of pharmaceutical tablets and polymers. Our studies have demonstrated that NIR imaging based on Compovision can solve several issues such as long acquisition times and relatively low sensitivity of detection. NIR imaging with Compovision is strongly expected to be applied not only to pharmaceutical tablet monitoring and polymer characterization but also to various applications such as those to food products, biomedical substances and organic and inorganic materials.     

Investigation of Novel Quantum Dots/Proteins/Cellulose Bioconjugate Using NSOM and Fluorescence

We investigated the engineered bioconjugate of cadmium selenide core/zinc sulfide shell, (CdSe)ZnS, quantum dots (QDs) with genetically modified proteins using fluorescence spectroscopy, near-field scanning optical microscopy (NSOM) and spectroscopy (NSOS). The protein polymer was allowed to self-assemble to the bacterial microcrystalline cellulose surface through the cellulosic binding domain. Results from the sample containing the QDs/protein/cellulose assemblies suggest that QDs were arrayed along the cellulose surface. The spectroscopic change of spectroscopic properties of the QDs upon bioconjugation, indicating the interaction among the immobilized QDs and between the constructed protein and QDs.