Site-Specific Detection of Free Radicals in Membranes Using an Amphiphilic Spin Trap

The detection of free radicals and related species has attracted considerable attention in recent years due to their critical roles in physiological and pathological processes. Among the various methods for the detection of free radicals, electron spin resonance coupled with spin-trapping technique has been an effective approach for the characterization and quantification of free radicals due to its high specificity. In this study, we designed and synthesized a novel amphiphilic spin trap, 2-(diethoxyphosphoryl)-2-heptadecanyl-3,4-dihydro-2H-pyrrole-1-oxide (DEPHdPO), from 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide with a long hydrocarbon chain at the C-5 position of the pyrroline ring, providing the amphiphilic character. The free-radical-trapping ability of DEPHdPO was evaluated by capturing hydroxyl radicals (^·OH), superoxide anions (\( {\text{O}}_{2}^{ \cdot - } \)), and carbon-centered free radicals in a model membrane prepared from sodium dodecyl sulfate (SDS). The results indicate that the hydrophobic hydrocarbon chain of DEPHdPO can be inserted into the inner core of SDS micelles, and the hydrophilic nitronyl functional moiety is located on the surface layer. Thus, various free radicals, including ^·OH radicals, \( {\text{O}}_{2}^{ \cdot - } \) anions, and carbon-centered radicals could be site-specifically detected near the membrane surface. Moreover, DEPHdPO could be successfully located on the surface of thylakoid membranes, and the nearby photo-initiated \( {\text{O}}_{2}^{ \cdot - } \) anions could be trapped site-specifically.     

荧光谱线分布对共焦荧光显微镜分辨率的影响

研究了荧光谱线分布对共焦荧光显微镜中分辨率的影响,导出了荧光谱线分布的荧光功率传输函数、三维脉冲响应函数.数值计算了荧光谱线均匀分布的情况,结果表明：与采用单一中心荧光波长的分辨率比较,共焦荧光显微镜的横向分辨率、纵向分辨率随着荧光谱线范围和色散的增大而下降;谱线响应范围小的探测器有利于分辨率.     

A Novel Fluorescent Biosensor for Detection of Silver Ions Based on Upconversion Nanoparticles

In this paper, we report a simple and sensitive fluorescent biosensor for the quantitative analysis of silver ions (Ag⁺) by using NaYF₄:Yb³⁺, Tm³⁺ upconversion nanoparticles (UCNPs). Ag⁺ could oxidize o-phenylenediamine (OPD) to the oxidized OPD (oxOPD) directly. The fluorescence of UCNPs can be significantly quenched by oxOPD through inner filter effects (IFE). Under the optimized conditions, the Ag⁺ concentration is proportional to the changes of the fluorescence intensity of UCNPs. The proposed method shows high selectivity and Ag⁺ could be quantitatively detected in the range of 0 to 0.5 mM with a low detection limit of 33 nM for Ag⁺. The selectivity and sensitivity of the detection can also be satisfactory. More importantly, this method has potential in practical application to detect Ag⁺ in real samples without interference.     

A Novel Fluorescent Probe for the Highly Selective and Sensitive Detection of Palladium in Aqueous Medium

Based on the Pd⁰-catalyzed Tsuji-Trost allylic oxidative insertion reaction, we developed a fluorescent probe PdL1 for sensing Pd⁰. As expected, probe PdL1 exhibited high selectivity and excellent sensitivity in both absorbance and fluorescence detection of Pd⁰ in CH₃CH₂OH/PBS (10 mM, pH = 7.4, 6:4, v/v) solution. The detection limit was calculated to be as low as 15 nM, which can meet the selective requirements for practical application.     

A Novel Single-Labeled Fluorescent Oligonucleotide Probe for Mercury(II) Ion Detection: Using the Inherent Quenching of Deoxyguanosines

A novel single-labeled fluorescent oligonucleotide (OND) probe for the detection of nanomolar mercury(II) ion in aqueous solution is developed based on the inherent quenching of deoxyguanosines. The formation of hairpin structure of OND-Hg²⁺ complex brings deoxyguanosines close to dye, resulting in decreased dye fluorescence due to photoinduced electron transfer from dye to deoxyguanosines.     

利用新型荧光银纳米团簇实现谷胱甘肽的快速精确检测

谷胱甘肽(GSH)是一种含有巯基的三肽分子,参与许多细胞内生化过程,具有抗氧化和整合解毒功能,在生物体内以及医学,食品等领域有着极为重要的作用。GSH参与细胞内、体液中的许多重要生化反应,其在人体内含量的变化,相应地提示了人体的健康问题。目前对GSH的检测手段有表面增强拉曼光谱(SERS)、电化学分析、高效液相色谱(HPLC)等,这些方法大都操作复杂、耗时较长或者需要昂贵的仪器。利用一种新型荧光银纳米团簇(Ag NCs)作为探针,通过同时分析银纳米团簇的荧光强度变化以及荧光峰位置移动实现了GSH的高精度快速检测。在检测过程中,GSH分子与荧光探针发生化学反应,改变了荧光探针的光化学特性,其荧光强度因发生猝灭而减弱,且其荧光峰位置因配体的改变也发生移动。通过对照组实验,我们进一步证明了所发展的检测方法对GSH目标具有很好的特异性,综合考察荧光强度和波长的变化数据可以很好地区分GSH以及其他结构类似的分子,同时探针对于多种盐离子及氨基酸等不敏感,能够很好地保证检测的准确性。我们报导的荧光探针合成步骤简单,过程绿色环保,GSH检测的响应速度快、光谱波动较小、相对误差小。进一步的研究有望实现细胞内的GSH高精度检测及成像。     

An Acid-Inert Fluorescent Probe for the Detection of Nitrite

The fluorescent detection of nitrite ions has been challenging owing to the poor acid-stability of typical fluorescent probes. Herein, an acid-inert rhodamine-based fluorescent probe has been developed for the fast detection of trace amount of nitrite ions with turn-on mode. The detection limit of nitrite ions was determined to be 9.4 nM (S/N = 3) and the linear range with high linear correlation was observed between 0.025 to 2.5 μM. A test paper method was developed for rapid visual detection of nitrite ions. Quantitative analysis of two real water samples also confirmed the reliability and sensitivity of the developed fluorescent method.     

A Schiff-Based Colorimetric Fluorescent Sensor with the Potential for Detection of Fluoride Ions

A simple Schiff-based colorimetric fluorescent receptor 1 was prepared. It exhibits a “turn-on-type” mode with high sensitivity in the presence of F^?. The change in color is very easily observed by the naked eye in the presence of F^?, whereas other anions do not induce such a change. Job plot indicated a 1:2 complexation stoichiometry between receptor 1 and F^?. The association constant for 1-F^? in CH₃CN was determined as 1.32*10⁵ M^?2 by a Hill plot.

新颖的酸催化迈克尔型巯基荧光探针的机理

迈克尔加成反应通常为碱催化的反应,但是在本工作体系当中,2-(2-乙烯基,7-羟基喹啉)丙二酸(QMA)能够被酸催化检测巯基. 从实验和理论计算证明了QMA探测巯基是,且溶液pH值越低,活 性越强. 在碱性溶液中,它的负离子形式QMA[-2H⁺]2^-不能够与电离前或电离后的巯基反应. 与QMA相比,它的酯QME显示出相反的pH效应,这与一般的基于迈克尔加成的巯基荧光探针类似. DFT计算得到的活化焓支持上述两种探针的pH效应. 在酸性溶液中,质子化的QMA与中性和碱性溶液中的负离子形式相比更容易与亲核试剂发生加成,因此能够与低活性的未电离硫醇反应. 相反地,QME与未电离硫醇反应的能垒很高,与电离后的硫醇反应活化能很低. 理论计算研究显示QMA与未电离硫醇的反应为1,4-加成而QME与电离后的硫醇的反应为1,2-加成.     

基于氧化石墨烯荧光适体传感器的多巴胺检测

以修饰有荧光基团(FAM)的多巴胺核酸适体作为识别元件,氧化石墨烯为猝灭剂,构建了光学适体传感器用于检测多巴胺。通过π-π堆积作用力,氧化石墨烯以共振方式把核酸适体上FAM能量转移到其表面,荧光信号消失;加入多巴胺后荧光恢复,荧光强度恢复的大小与多巴胺浓度呈正相关关系。实验优化结果表明,在反应时间5 min和10μg/mL氧化石墨烯条件下,氧化石墨烯可以达到对FAM的最高猝灭效率;25min孵育后,多巴胺恢复荧光强度达到稳定;传感器线性检测范围为1~500μmol/L,检测限达到1μmol/L。所制备传感器具有检测范围宽、检测速度快、特异性强以及检测成本低等优点。     

Polar-Core Spin Vortex of Quasi-2D Spin-2 Condensate in a Flat-Bottomed Optical Trap

Motivated by the recent experiments realized in a flat-bottomed optical trap [Science 347(2015) 167;Nat. Commun. 6(2015) 6162], we study the ground state of polar-core spin vortex of quasi-2D spin-2 condensate in a homogeneous trap plus a weak magnetic field. The exact spatial distribution of local spin is obtained and the vortex core are observed to decrease with the growth of the effective spin-spin interaction. For the larger effective spin-spin interaction, the spatial distribution of spin magnitude in spin-2 condensate we obtained agrees well with that of spin-1 condensate in a homogeneous trap, where a polar-core spin vortex was schematically demonstrated as a fully-magnetized planar spin texture with a zero-spin core. The effective spin-spin interaction is proportional to both the bare spin-spin interaction and the radius of the homogeneous trap, simultaneously. Thus the polar-core spin vortex we obtained can be easily controlled by the radius of the trap.     

A Novel Fluorescent Nanoparticle for Sensitive Detection of Cry1Ab Protein In Vitro and In Vivo

Here, we report the synthesis and characterization of CoFe₂O₄ doping Ag₂S dendrimer-modified nanoparticles (CoFe₂O₄-Ag₂S DMNs) in Cry1Ab protein detection and imaging. The near-infrared Ag₂S quantum dots were first prepared by using the thermal decomposition method, followed by modification of the water-soluble quantum dots using the method of solvent evaporation and ligand exchange, and finally the fluorescent magnetic bifunctional nanoparticles were obtained by binding with CoFe₂O₄. As-prepared CoFe₂O₄-Ag₂S DMNs were characterized by fluorescence (FL) spectroscopy and transmission electron microscopy (TEM). Results showed that Ag₂S DMNs could sensitively detect Cry1Ab both in vitro and in vivo. In vitro, the enhanced FL intensity as a function of the concentration is notably consistent with the Langmuir binding isotherm equation in the range of 0–200 ng/mL of Cry1Ab proteins. The detection limit of this method was found to be 0.2 ng/mL. Meanwhile, the fluorescence wavelength was extended to the second near-infrared range (NIR-II, 1.0~1.4 μm), which enables in vivo imaging. This study highlights the importance of NIR QDs doping magnetic materials as a new method to trace Bacillus thuringiensis (Bt) in insects and their potential applications in in vivo NIR tissue imaging.     

An Improvement on the Combination of Magnetic Trap and Fluorescent Resonant Energy Transfer

The combination of magnetic trap(MT) and fluorescence resonant energy transfer(FRET) allows for nanoscale measurements of configurational changes of biomolecules under force. However, the magnetic bead involved in MT experiments introduces a substantial amount of background fluorescence which reduces the signal-to-noise ratio(SNR) of FRET significantly. Moreover, the short lifetime of the dye used in FRET limits the total sampling time when combined with MT. Here we use a moveable tube lens to adjust the wave front in the light pathway of MT so that both images of the magnetic bead and the fluorescent signals can be detected when long DNA handles are used to reduce the auto-fluorescence of the magnetic bead. We utilize the internal trigger of an electron multiplying charge-coupled device camera to control a shutter so that the dye can be excited intermittently when long time measurement of FRET is needed. As a demonstration of the hybrid technique, we observe the unfolding/refolding dynamics of a DNA hairpin and measure the DNA unwinding activity of the saccharomyces cerevisiae Pif1(Pif1). Our results show that the unwinding burst of Pif1 under external force is different from that without the force. In addition, the improvement provides a better SNR and a longer sampling time in experiments in the MT-FRET assay.     

A Hybrid Coumarin-Semifluorescein-Based Fluorescent Probe for the Detection of Cysteine

A new type of turn-on fluorescent probe CF-AC for the detection of Cys was firstly reported. The probe exhibited an excellent response to Cys with high selectively and sensitivity. In the presence of Cys, two fluorescence emission peaks at 525 nm and 650 nm appeared accompanied by the fluorescence color change from blue to red. Morever, the probe had good biocompatibility and could be successfully used for fluorescence imaging of Cys in MCF-7 cells.     

Study of Polycation-Capped Mn:ZnSe Quantum Dots as a Novel Fluorescent Probe for Living Cells

Transition metal manganese ion (Mn²⁺) doped zinc selenide quantum dots (Mn:ZnSe D-Dots) have been considered as a new material for fluorescent probes in biological labeling. However, this application is limited by the low membrane permeability of D-Dots. In this work, Mn:ZnSe D-Dots were capped with the polycation Sofast to label living cells. For the first time, the efficiency of cellular uptake in living cells is significantly enhanced. Various molar ratios of Sofast to D-Dots were explored and compared to obtain the optimal reaction conditions between Sofast and D-Dots for preparing Sofast/D-Dots nano-compound. A comparison on the fluorescence labeling ability of living cells were made between Sofast/D-Dots and pure D-Dots. Results from laser scanning confocal microscope show that Sofast/D-Dots complexes enter the cells more efficiently than pure D-Dots, even with a lower concentration and shorter incubation time. The cytotoxicities of D-Dots and Sofast/D-Dots were also studied. It was found that Sofast/D-Dots have a much lower cytotoxicity than cadmium-containing quantum dots (i.e. CdTe and CdTe/ZnS). Our results suggest that the non-heavy-metal-containing Sofast/D-Dots complexes have a great potential in the application of biological labeling, especially of long-time bioimaging in living cells.     

Analytical Solutions for the Two-Dimensional Gross-Pitaevskii Equation with a Harmonic Trap

Both the homotopy analysis method and Galerkin spectral method are applied to find the analytical solutions of the two-dimensional and time-independent Gross-Pitaevskii equation,a nonlinear Schrdinger equation used in describing the system of Bose-Einstein condensates trapped in a harmonic potential.The approximate analytical solutions are obtained successfully.Comparisons between the analytical solutions and the numerical solutions have been made.The results indicate that they are agreement very well with each other when the atomic interaction is not too strong.     

光栅单色仪最佳分度的判断

本文利用波动光学的简单方法讨论光栅单色仪的分辨极限与步进电机参数的联系,为最佳利用分辨极限提供了简单可行的实验方法,也给出了用步栅单色仪的分辨率可供参考的例证。     

A BODIPY Based Fluorescent Probe for the Rapid Detection of Hypochlorite

A new boron-dipyrromethene (BODIPY) fluorescent dye aimed at sensitively detecting hypochlorite anion (ClO^?) has been designed, synthesized and characterized. The probe is comprised of a BODIPY fluorophore unit and a ClO^? specific reactive group of amidoxime. The addition of hypochlorite results in a red-shift of absorption and emission spectra of the probe accompanied by a decrease of intensity and spectra changes (A₅₀₀ and 1/I₅₁₂) of the probe can achieve a good linearity to the concentration of ClO^?. The fluorescence probe can react to ClO^? rapidly (within 60 s) in a wide pH range (4–10) with high sensitivity (detection limit of 6.81 μM) and selectivity. The reaction mechanism has been proposed and confirmed by MS analysis, ClO^? anion oxidizes amidoxime moiety to hydroxyl group and hydroxyl group is further oxidized to formyl group in the formation of a corresponding aldehyde compound. In addition, the probe has also been successfully applied to detect ClO^? in tap water and river water samples by spiking a known amount of standard ClO^?.