Fluorescence Imaging of Stem Cells,Cancer Cells and Semi-Thin Sections of Tissues using Silica-Coated CdSe Quantum Dots

Trioctylphosphine oxide capped cadmium selenide quantum dots, synthesized in organic media were rendered water soluble by silica overcoating. Silanisation was done by a simple reverse microemulsion method using aminopropyl silane as the silica precursor. Further, the strong photoluminescence of the silica-coated CdSe quantum dots has been utilized to visualize rabbit adipose tissue-derived mesenchymal stem cells (RADMSCs) and Daltons lymphoma ascites (DLA) cancerous cells in vitro. Subsequently the in vivo fluorescence behaviours of QDs in the tissues were also demonstrated by intravenous administration of the QDs in Swiss albino mice. The fluorescence microscopic images in the stem cells, cancer cells and semi-thin sections of mice organs proved the strong luminescence property of silica-coated quantum dots under biological systems. These results establish silica-coated CdSe QDs as extremely useful tools for molecular imaging and cell tracking to study the cell division and metastasis of cancer and other diseases.     

Silica-coated quantum dots fluorescent spheres synthesized using a quaternary ‘water-in-oil’ microemulsion system

Nanoscale and microscale silica spheres embedded with multiple CdSe quantum dots (QDs, having average diameters of about 2.4 and 5.0 nm, respectively.) were synthesized by using a quaternary ‘water-in-oil’ microemulsion. Comparing the uncoated QDs, the quantum yields (QYs) of the silica-coated QD spheres were enhanced when the QD cores were synthesized using mercaptoacetic acid (MA) as a stabilizer, while the QYs were dramatically decreased when the cores were synthesized using citric acid (CA) as a stabilizer. The enhanced QYs could be further improved by heating the silica-coated QDs in aqueous solution. Although the QYs of the silica-coated QDs were not high, these spheres emitted bright fluorescence. The silica shells contained numerous micropores (∼0.58–0.91 nm), and small amounts of toxic ions (such as Cd²⁺) could be released from the silica spheres. However, the release rate of toxic ions from the silica spheres was significantly reduced compared with that of the uncoated QDs.     

Improving colloidal properties of quantum dots with combined silica and polymer coatings for in vitro immuofluorenscence assay

Semiconductor quantum dots (QDs) are promising fluorescence probes for immuofluorescence assay in the biological applications. However, water solubilization and non-specific binding are two critical issues to be addressed for the practical uses. Here, we reported a new type of QDs with combined silica and polymer coating. QDs with excellent colloidal properties were prepared via carboxylation of the amino groups on the surface of silica-coated QDs by reacting with multi-carboxyl poly (acrylic acid) (PAA). Hydrodynamic size of PAA-functionalized silica-coated QDs was around 40 nm. They were highly fluorescent (about 47.8% quantum yield). No precipitate of QDs was observed after 3 month storage at 4 °C. When cancer cells (HeLa) were used, the functionalized QDs exhibited little or no non-specific cellular binding. The results from in vitro experiments indicated that PAA-functionalized silica-coated QDs-antibody bioconjugates had excellent antigen-capture ability and exhibited little or no non-specific binding to polystyrene spheres which were used to immobilize the antigen for immuoflurescence assay. The PAA-functionalized silica-coated QDs with improved colloidal properties could serve as excellent alternative fluorescent probes for biodetection.     

Synthesis,biocompatibility and luminescence properties of quantum dots conjugated with amino acid-functionalized β-cyclodextrin

A series of CdSe and CdSe/CdS quantum dots (QDs) labeled with amino acid-modified β-cyclodextrin (β-CD) was prepared by a simple ultrasonic method. These amino acid-modified β-CD-coated QDs are very soluble and stable in biological buffer. They also have high colloidal stability and strong optical emission properties that are similar to those of untreated tri-n-octylphosphine oxide (TOPO)-coated QDs. The quantum yields (QYs) of these amino acid-modified β-CD-coated CdSe and CdSe/CdS QDs in biological buffer were found to be very high. In particular, the QYs of the positively charged l-His-β-CD-coated CdSe/CdS QDs were as high as 33.5±1.8%. In addition, the fluorescence lifetime of these QDs was also very long in PBS solutions as determined by frequency domain spectroscopy. For example, the lifetime of l-His-β-CD-coated CdSe/CdS QDs was 8.6 ns. The in vitro cytotoxicity of these QDs in ECV-304, SH-SY5Y and HeLa cells was found to be lower. l-His-β-CD-coated CdSe/CdS QDs were the least cytotoxic (IC₅₀ 95.6±3.2 mg mL^?1 in ECV-304 cells after 48 h). The flow cytometry results show that the positively charged amino acid led to a considerable increase in biocompatibility of QDs. This may be attributed to the presence of an amino acid-modified β-CD outer layer, which enhanced the biocompatibility.     

Photostability comparison of CdTe and CdSe/CdS/ZnS quantum dots in living cells under single and two-photon excitations

The photostability is an outstanding feature of quantum dots (QDs) used as fluorescence probes in biological staining and cell imaging. To find out the related factors in the QD photostability, the photobleaching of naked CdTe QDs and BSA coated CdSe/CdS/ZnS QDs in human hepatocellular carcinoma (QGY) cells and human nasopharynx carcinoma (KB) cells were studied under single photon excitation (SPE) and two-photon excitation (TPE). In these two cell lines the cellular QDs were irradiated by a 405 nm continuous wave laser for SPE or an 800 nm femto-second (fs) laser for TPE. The QD photobleaching with the irradiation time was found to fit a biexponential decay. The fast decay plays a dominant role in the bleaching course and thus can be used as the parameter to quantitatively evaluate the QD photostability. The TPE decreased the QD photobleaching as compared to SPE. The BSA coated core/shell QDs had improved the photostability up to 4-5 times than the naked QDs due to the shielding effect of the QD shell. Therefore, it is better to use core/shell structured QDs as the fluorescence probe combining with a TPE manner for those long-term monitoring studies.     

A Dual-Functional Ferroferric Oxide/Quantum Dots Theranostic Nanoplatform for Fluorescent Labeling and Photothermal Therapy

A biocompatible, nontoxic theranostic nanoplatform consisting of mesoporous silica-coated ferroferric oxide (Fe₃O₄) and Mn-doped ZnS-ZnS quantum dots (QDs) is synthesized via a layer-by-layer method. Transmission electron microscopy, X-ray diffractometer, magnetometry, and fluorophotometer are employed to characterize the nanoplatform. The nanoplatform exhibits excellent superparamagnetic, fluorescent, and light absorption properties. The template method is introduced to form a mesoporous silica structure on the nanoplatform, lowering the mass of the nanoplatform and effectively promoting the absorption efficiency of the incident light compared with the traditional silica layer. In addition, after endocytosis of the nanoplatform, cancer cells are easily detected under a fluorescence microscope because of the excellent fluorescent behavior of QDs. Moreover, in vitro experiments confirm that nanoplatform possesses perfect photothermal effect to destroy tumor cells under laser irradiation. Therefore, ferroferric oxide/QDs nanoplatforms, combined with the functions of fluorescent labeling and photothermal therapy for cancer cells, are expected to be a promising biopotential material in the field of diagnosis and treatment.     

Non-specific interactions of CdTe/Cds Quantum Dots with human blood mononuclear cells

In order to study biological events, researchers commonly use methods based on fluorescence. These techniques generally use fluorescent probes, commonly small organic molecules or fluorescent proteins. However, these probes still present some drawbacks, limiting the detection. Semiconductor nanocrystals - Quantum Dots (QDs) - have emerged as an alternative tool to conventional fluorescent dyes in biological detection due to its topping properties - wide absorption cross section, brightness and high photostability. Some questions have emerged about the use of QDs for biological applications. Here, we use optical tools to study non-specific interactions between aqueous synthesized QDs and peripheral blood mononuclear cells. By fluorescence microscopy we observed that bare QDs can label cell membrane in live cells and also label intracellular compartments in artificially permeabilized cells, indicating that non-specific labeling of sub-structures inside the cells must be considered when investigating an internal target by specific conjugation. Since fluorescence microscopy and flow cytometry are complementary techniques (fluorescence microscopy provides a morphological image of a few samples and flow cytometry is a powerful technique to quantify biological events in a large number of cells), in this work we also used flow cytometry to investigate non-specific labeling. Moreover, by using optical tweezers, we observed that, after QDs incubation, zeta potentials in live cells changed to a less negative value, which may indicate that oxidative adverse effects were caused by QDs to the cells.     

异丙醇对油溶性CdSe团簇量子点荧光的影响

为了探究异丙醇对胶体CdSe团簇量子点荧光光谱的影响,使用胶体化学法在油酸-石蜡体系下合成CdSe团簇量子点,对团簇量子点和异丙醇采用不同的混合比例,得到了其修饰之后的荧光发光谱(PL)和紫外吸收(UV)光谱,并采用曲线拟合的方法对团簇量子点的光学性能进行了研究。通过对AFM和傅里叶红外光谱的分析,探讨了CdSe团簇量子点光学性能改变的内在联系。结果表明:随着异丙醇浓度的增加,量子点的荧光峰出现11 nm的蓝移,且蓝移曲线呈阶梯状变化;相对荧光强度也呈现出先上升再下降的波动性变化,且波动幅度最大能达到1 000 a.u.;量子点的第一吸收峰和第二吸收峰都会发生不同程度的红移,且最大红移达到12 nm。     

复合荧光CdSe量子点-脂质体的制备与表征

通过脂质体方法成功地将三辛基氧化膦(TOPO)包覆的CdSe发光量子点从非极性有机溶剂转移到生物相容性的水溶液中.分别通过透射电镜(TEM)、荧光Mapping图像,以及光致发光(PL)光谱进行表征.TEM照片显示制备的CdSe核量子点为球形,具有良好的单分散特性,平均粒径约为3nm.CdSe-脂质体复合体的平均尺寸大约20nm,TEM清楚地显示了CdSe量子点被诱捕在脂质体中.荧光Mapping显示了CdSe-脂质体复合体的发光强度分布.脂质体方法转移TOPO包覆的CdSe量子点,借助了磷脂的双分子链与CdSe表面的TOPO配体之间的疏水相互作用,在CdSe的第一配体层外部形成第二配体层,保留了CdSe的存在环境,光致发光光谱表明,量子点-脂质复合体基本保持了CdSe核量子点的发射效率.     

Encapsulation of highly confined CdSe quantum dots for defect free luminescence and improved stability

This is the first report on the generation of trap states and their effective elimination in highly confined CdSe quantum dots in order to obtain enhanced and stable optical properties prepared by aqueous route. Surface plays an important role in optical properties of quantum dots (QDs) and surface modification of quantum dots can improve optical properties. In present work luminescent CdSe QDs were prepared using 2-Mercaptoethanol (2-ME) as stabilizing agent and encapsulated by polymer. Different concentrations of 2-ME were used to tune the emission spectra with respect to their reduced size. Addition of 2-ME to CdSe QDs enhances the trap emission and quenching band edge emission due to (i) increased surface to volume ratio and; (ii) presence of high concentration of sulfide ions as confirmed from EDX analysis as sulfide ions possesses the hole scavenging characteristics. Polymer encapsulation of QDs was carried out to make them stable and to improve their optical properties. Even though there are previous reports addressing the improved optical properties by polymer encapsulation and silica encapsulation but experimentally it has not been reported yet experimentally. In this work we have synthesized and characterized water soluble polymer encapsulated QDs and proved the facts experimentally. Photoluminescence spectroscopy clearly reveals the role of polymer encapsulation in boosting the optical properties of CdSe QDs. FTIR spectra validate the presence of biocompatible functional groups on CdSe4/PEG (Polymer encapsulated QDs).     

CdSe/ZnSe/ZnS多壳层结构量子点的制备与表征

展示了一种简捷的多壳层量子点合成路线。在含有过量Se源的CdSe体系中直接注入Zn源,"一步法"合成了CdSe/ZnSe量子点;进一步以CdSe/ZnSe为"核",表面外延生长ZnS壳层制备了核/壳/壳结构CdSe/ZnSe/ZnS量子点。相对于以往报道的多壳层结构量子点的制备方法,该方法通过减少壳层的生长步骤有效地简化了实验操作,缩短了实验周期,同时减少对原料的损耗。对量子点进行高温退火处理,能够大幅提高CdSe/ZnSe/ZnS量子点的发光量子产率。透射电镜、XRD以及光谱研究表明:所制备的量子点接近球形,核与壳层纳米晶均为闪锌矿结构,最终获得的CdSe/ZnSe/ZnS量子点的光致发光量子产率达到53％。为了实现量子点的表面生物功能化,通过巯基酸进行了表面配体交换修饰,使量子点表面具有水溶性的羧基功能团,并且能够维持较高的光致发光量子产率。     

Synthesis of CdSe/CdS with a simple non-TOP-based route

A convenient and non-TOP-based route for the synthesis of core-shell CdSe/CdS quantum dots (QDs) is developed for the first time. Simple reagents, such as cadmium oxide, selenium powder, sodium sulfide, paraffin and oleic acid with obvious advantages are used to replace organometallics. This simple route allows the preparation of a series of core-shell CdSe/CdS QDs emitting in a wide wavelength range (from 510 to 615 nm). After passivation of CdSe by CdS shell using sodium sulfide as the source of sulfur at 80 °C, the quantum yields (QYs) are improved from 15-30% to 35-50% and remained stable at least for 4 months. A narrow bandwidth (FWHM<50 nm) indicates that the as-prepared QDs have uniform size distribution, desirable dispersibility and good fluorescence properties. The whole procedure can be carried out either open to air or under nitrogen atmosphere, which is simpler, greener and cheaper as compared with TOP-based route.     

脂溶性CdSe/ZnS量子点脂质体复合物的制备及表征

在油相中成功合成了脂溶性CdSe/ZnS核壳量子点纳米粒,粒径平均为4.5 nm,量子产率达29%,发射波长为540 nm。通过薄膜分散法,以蛋黄卵磷脂、胆固醇为膜材,将脂溶性的CdSe/ZnS核壳量子点包覆于脂质体磷脂双分子层中,由于磷脂分子的两亲性,使得脂溶性的CdSe/ZnS核壳量子点同时又具有亲水性。通过透射电镜对脂质体形态进行了表征,倒置荧光显微镜证实了发光CdSe/ZnS核壳量子点成功包埋于脂质体双分子层中,包裹的发光CdSe/ZnS核壳量子点具有更稳定的发光及抗光漂白性质。     

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.     

Phase transfer of agglomerated nanoparticles: deagglomeration by adsorbing grafted molecules and colloidal stability in polymer solutions

We aimed to develop liposomes for loading both cisplatin and quantum dots (QDs) for both drug delivery and bioimaging. The resultant quantum-dot-liposomes (QDLs) with cisplatin were characterized using dynamic light scattering, transmission electron microscopy (TEM), encapsulation efficiencies, and fluorescence intensity. QDLs composed of CdSe or CdSe/ZnS QDs represented a size of about 100?nm. The QDLs were prepared at a high QD loading efficiency of nearly 100?%. Most QDs were located within the liposomal bilayers as evidenced by TEM. Slow and sustained cisplatin release from QDLs was achieved. The cellular uptake of QDLs demonstrated effective internalization and significant fluorescence in melanoma cells. The signal derived from QDLs could be observed by different wavelength settings. The cisplatin-containing QDLs revealed higher cytotoxic activity compared to an equal dose of free cisplatin. CdSe/ZnS QDLs were intravenously administered to mice, and the biodistribution was observed with an in vivo imaging system. Significant fluorescence signal and cisplatin accumulation were detected in the brain and skin, as verified by ex vivo imaging and drug distribution. Liposomal inclusion could reduce the reticuloendothelial system uptake of QDs and cisplatin. QDLs evaluated in this study represent a new potential method for theranostic purposes.     

Two-step annealed CdS/CdSe co-sensitizers for quantum dot-sensitized solar cells

CdS/CdSe co-sensitizers on TiO₂ films were annealed using a two-step procedure; high temperature (300 °C) annealing of TiO₂/CdS quantum dots (QDs), followed by low temperature (150 °C) annealing after the deposition of CdSe QDs on the TiO₂/CdS. For comparison, two types of films were prepared; CdS/CdSe-assembled TiO₂ films conventionally annealed at a single temperature (150 or 300 °C) and non-annealed films. The 300 °C-annealed TiO₂/CdS/CdSe showed severe coalescence of CdSe QDs, leading to the blocked pores and hindered ion transport. The QD-sensitized solar cell (QD-SSC) with the 150 °C-annealed TiO₂/CdS/CdSe exhibited better overall energy conversion efficiency than that with the non-annealed TiO₂/CdS/CdSe because the CdSe QDs annealed at a suitable temperature (150 °C) provided better light absorption over long wavelengths without the hindered ion transport. The QD-SSC using the two-step annealed TiO₂/CdS/CdSe increased the cell efficiency further, compared to the QD-SSC with the 150 °C-annealed TiO₂/CdS/CdSe. This is because the 300 °C-annealed, highly crystalline CdS in the two-step annealed TiO₂/CdS/CdSe improved electron transport through CdS, leading to a significantly hindered recombination rate.     

CdSe量子点的光物理性质研究与荧光杂化纤维的制备

半导体纳米晶体（NCs）具有良好的光稳定性,广泛的发射持久性和高消光系数,在过去几年被广泛研究报道,其中,硒化镉半导体纳米晶体(CdSe NCs)被广泛用于电子照明、太阳能发电、光电传感等领域。然而CdSe NCs的电学、热力学和光物理性质具有较强的尺寸依赖性,在传统的制备方法及应用中容易出现晶体表面缺陷和悬空键以及较为严重的生物毒性和环境毒性。为了实现量子点在各个领域的应用,必须严格控制CdSe NCs的发光波长,尺寸分布以及荧光性能。本研究通过高温热注射法合成了单分散的胶体发光CdSe NCs,使用表面配体对CdSe NCs进行修饰,研究了不同烷基链长度的配体对CdSe NCs尺寸分布和荧光性能的影响,并通过改变溶剂配比制备了纺丝溶液,将其与聚乙烯吡咯烷酮（PVP）进行杂化,制备了PVP/CdSe QDs荧光杂化纤维。结果表明,与传统CdSe NCs相比,经表面配体的修饰的CdSe NCs在有机溶液中因分子间吸附的降低在溶液中有良好的稳定性,具有可调节的溶解度,弥补了缺陷和悬空键造成的荧光性能下降。在CdSe晶体结构的形成过程中,表面配体也有着显著的调控作用。并且更为重要的是,该研究将表面配体修饰与杂化相结合,改善了表面配体的附着,在杂化材料的制备过程中避免了硒化镉纳米晶体与高分子基体直接接触,为荧光团提供了良好的发光微环境,保证了CdSe NCs的荧光性能,使杂化纤维具有良好而稳定的荧光性能。同时,PVP的引入使CdSe NCs的生物毒性和环境毒性得以改善,使材料更加环境友好且具有更好的生物相容性,大大提升了材料的应用范围。事实证明,PVP/CdSe QDs杂化微纤维杂化相容性和分散性良好,具有优异的荧光性能和材料成型性,纤维合成方式简便易行且造价低廉,可应用于溶液处理,光学照明,电极材料,和生物成像等各个领域。     

Demonstration of a nanoparticle-based optical diode

We present the operation of an optical device that exhibits diodelike properties based on two adjacent layers of quantum dots (QDs) encased in a fiber-optic jacket. The possibility of a multilayered device is also discussed. A significant change in the emission spectrum of CdSe/ZnS core-shell QDs was observed when excited by the input laser and the fluorescence of other CdSe/ZnS core-shell QDs. The output of the diode can be taken to be either the incoming laser wavelength of light similar to a conventional diode, or the output may be considered to be one of the QD fluorescence wavelengths. Current work has applications in biological fluorescence monitors and sensors as well as in telecommunications applications.     

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.     

CdSe quantum dots decorated by mercaptosuccinic acid as fluorescence probe for Cu

Water-soluble CdSe quantum dots (QDs) were synthesized using mercaptosuccinic acid (MSA) as a stabilizer. The growth process and characterization of CdSe quantum dots were determined by transmission electron microscopy (TEM), X-ray diffraction (XRD), photoluminescence (PL) spectroscopy, Ultraviolet-visible (UV-vis) spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy. Results demonstrated the MSA-capped CdSe QDs were highly crystalline and possessed good optical properties. Further, the resulting products could be used as fluorescent probes to detect Cu²⁺ ions in physiological buffer solution. The response was linearly proportional to the concentration of Cu²⁺ ion in the range 2×10⁻⁸- 3.5×10⁻⁷ mol L⁻¹ with a detection limit of 3.4 nmol L⁻¹.