Recent Development of DNA-modified AIEgen Probes for Biomedical Application

A variety of DNA-based probes are utilized for the detections of multiple analytes and DNA nanotechnology has been thriving for recent decades and achieving numerous nanostructures,mainly focusing on DNA morphology modulation and multifunctional systems engineered into to the complicated works.Among the numerous detections,fluorescence method is a non-invasive,highly selective and sensitive means for varieties of applications,but their emissions are often compromised by the aggregation-caused quenching(ACQ)effect,which weakens their applications.The aggregation induced emission luminogens(AIEgens)are created with non emissive or weakly emissive in a low concentration but emit strong fluorescence in a high concentration with aggregated states.Herein,numerous functionalized AIEgens have been emerged and used for detection and imaging and DNA-modified AIEgen probes are introduced.In this vein,here we report the progress on DNA-modified AIEgen probes in recent years and highlight their conjugation strategies including covalent bonding,electrostatic interaction and their applications of biosensing.Moreover,multiple DNA strands are needed to introduce into the DNA-modified AIEgen probes for more purposes.At the end,some challenges are mentioned to discuss the new trend of DNA-modified AIEgen probes.     

High-Affinity Ratiometric Fluorescence Probe Based on 6-Amino-2,2′-Bipyridine Scaffold for Endogenous Zn2+ and Its Application to Living Cells

Zinc is an essential trace element involved in many biological activities; however, its functions are not fully understood. To elucidate the role of endogenous labile Zn²⁺, we developed a novel ratiometric fluorescence probe, 5-(4-methoxyphenyl)-4-(methylsulfanyl)-[2,2′-bipyridin]-6-amine (6 (rBpyZ)) based on the 6-amino-2,2′-bipyridine scaffold, which acts as both the chelating agent for Zn²⁺ and the fluorescent moiety. The methoxy group acted as an electron donor, enabling the intramolecular charge transfer state of 6 (rBpyZ), and a ratiometric fluorescence response consisting of a decrease at the emission wavelength of 438 nm and a corresponding increase at the emission wavelength of 465 nm was observed. The ratiometric probe 6 (rBpyZ) exhibited a nanomolar-level dissociation constant (K_d = 0.77 nM), a large Stokes shift (139 nm), and an excellent detection limit (0.10 nM) under physiological conditions. Moreover, fluorescence imaging using A549 human lung adenocarcinoma cells revealed that 6 (rBpyZ) had good cell membrane permeability and could clearly visualize endogenous labile Zn²⁺. These results suggest that the ratiometric fluorescence probe 6 (rBpyZ) has considerable potential as a valuable tool for understanding the role of Zn²⁺ in living systems.     

发夹型荧光分子探针用于DNA甲基化酶的活性分析及其在药物筛选中的应用

报道了一种基于发夹型荧光探针的甲基化酶活性的分析方法, 甲基化酶和相应的限制性内切酶的识别位点被设计在发夹型探针的茎部, 四甲基罗丹明(TAMRA)被连接在探针的5'端, 其荧光被连在3'端的熄灭基团4-(4'-二甲基对胺基偶氮苯)苯甲酸(DABCYL)所熄灭. 限制性内切酶可切割未发生甲基化修饰的探针, 导致探针的发夹结构遭到破坏, 引起TAMRA荧光信号的恢复. 根据荧光信号的恢复程度可实现对甲基化酶活性的分析. 在此基础上, 建立了一种简便、快速分析抗肿瘤药物对DNA甲基化酶活性的影响的方法, 为筛选针对基因甲基化异常引起的恶性肿瘤的治疗药物提供了一种新的思路和方法.     

Small molecule diffusion in a rubbery polymer near Tg: Effects of probe size,shape, and flexibility

A novel experimental approach involving fluorescence nonradiative energy transfer (NRET) is employed to study the Fickian diffusion of small molecules in rubbery poly(isobutyl methacrylate) (PiBMA) films near the glass transition, using a formalism that directly relates the small molecule translational diffusion coefficient, D, to changes in the normalized nonradiative energy transfer efficiency, E_N. Values of D for pyrene, 1,3-bis-(1-pyrene) propane (BPP), 1,3-bis-(1-pyrene) decane (BPD), 9,10-bis-phenyl ethynyl anthracene (BPEA), diphenyl Disperse Red 4 (DPDR4), and decacyclene in PiBMA are measured over temperatures ranging from approximately T_g to T_g + 25°C. Among these chromophores, significant differences in both the magnitude and temperature dependence of D are observed which are attributed to differences in molecule shape and flexibility, as well as molar volume. Other factors being equal, chromophore flexibility was shown both to increase the magnitude of D and to decrease its dependence on temperature, as does an increase in aspect ratio. For BPD, these effects are attributed to the ability of the flexible molecule to diffuse in a piecewise manner, requiring the cooperative mobility of fewer polymer chain segments than a rigid molecule of the same molar volume. For BPEA and DPDR4, this deviation from D being dominated by molar volume effects is attributed the to high aspect ratio of these elongated molecules. © 1996 John Wiley & Sons, Inc.     

Comparison of electron spin resonance probe and dielectric relaxation in physically and chemically crosslinked polymers

Dielectric relaxation spectroscopy in the frequency range from 10⁻² Hz to 10⁶ Hz and electron spin resonance (ESR) experiments are employed to study the dynamics in chemically and physically crosslinked networks. As examples for physically crosslinked networks ortho- and para-cresol novolacs were investigated. Dielectrically these materials show low-temperature β- and high-temperature α-relaxation. Both relaxation regions differ for both types of novolacs. This is also reflected by the ESR measurements and is discussed in terms of different hydrogen bonds found to be stronger in para-cresol novolac. For the chemically crosslinked poly(triallyl isocyanurate) only a β-peak is found by the dielectric measurements. Also in the ESR experiment the slow motion regime is characterized up to high temperatures. This means that the segmental motion is strongly suppressed by chemical crosslinking. Nevertheless the obtained change in the formal T_50G value can be used to characterize the glass transition in highly crosslinked systems by the ESR method.     

靶向分子羧甲基多糖与DNA作用机理研究

以溴化乙锭(EB)为荧光探针,结合紫外光谱法研究了壳聚糖、羧甲基壳聚糖、羧甲基纤维素与DNA的相互作用,药物分子与DNA作用程度可用作用常数D来表示,实验表明: 几种生物多糖与DNA分子的作用强弱顺序为: 壳聚糖＞羧甲基壳聚糖＞羧甲基纤维素;多糖主要以嵌插方式与DNA作用, 使EB-DNA荧光猝灭。     

荧光法研究牛血清白蛋白与三羟基苯基荧光酮-钼(Ⅵ) 配合物探针的作用机理

用荧光光谱法研究了三羟基苯基荧光酮(TH-PF)-钼(Ⅵ)配合物与牛血清白蛋白的结合反应。探讨了TH-PF-Mo(Ⅵ)配合物对蛋白质内源荧光的猝灭机理,并测定了不同温度下的结合常数,温度为25 ℃时,荧光猝灭法测得该反应的结合常数为K=4.78×104 L·mol-1,温度为40 ℃时,荧光猝灭法测得该反应的结合常数为K=3.72×104 L·mol-1。根据Frster非辐射能量转移理论,确定了给体-受体之间的作用距离和能量转移效率(E=0.314),并根据热力学参数确定了TH-PF-Mo(Ⅵ)配合物与牛血清白蛋白之间的作用力类型,以静电引力为主。     

碳量子点/曙红Y比率型荧光探针测定L-抗坏血酸

以天然生物质去皮的蓖麻为碳源,采用一步水热法合成了荧光性能优良的绿色荧光蓖麻碳量子点(CO-CQDs),对其形貌和发光性能进行了表征。通过将该CO-CQDs与荧光极强的卤代荧光素染料曙红Y(EY)复合,二者可形成荧光发射峰相距较远的新型CO-CQDs/EY复合物。在pH=4.00的Na2HPO4-柠檬酸缓冲溶液中,在320 nm的激发波长下,CO-CQDs/EY复合物于405 nm和540 nm处显示出两个独立的荧光发射峰。在该体系中加入Cr(Ⅵ),405 nm和540 nm两处的荧光信号均显著猝灭。L-抗坏血酸(L-ascorbic acid,AA)的加入可使复合物于540 nm的荧光信号恢复,而405 nm处的荧光强度基本不变。据此建立了一种以CO-CQDs/EY复合物为比率型荧光探针测定AA的新方法。实验测定了荧光信号恢复的最佳条件和影响荧光恢复的因素,初步探讨了反应机理。在优化的实验条件下,该探针于540 nm/405 nm两处的荧光强度比值与AA的浓度在5.0×10^-8~4.0×10^-6 mol/L范围内呈良好线性关系,检出限为3.7×10^-8 mol/L。该探针用于检测药物、水果和蔬菜中AA的含量,结果满意。     

Imaging and tracking an electrostatic charge micro-domain by Kelvin force microscopy as evidence of water adsorption on mica surface

Muscovite mica is a widely accepted substrate for scanning probe microscopy (SPM) investigations. However, mica has intrinsic properties that alter samples and obstruct their analysis due to free charges build-up, ionic exchange and water adsorption taking place at the surface. In addition to interfacial phenomena, there is a growing interest in electrostatic charges on insulators as they are crucial in diverse applications. Despite the high demand for studies of this nature, experimental set-ups capable of resolving charge build-up at the micro-scale are still scarce and technically limited. Here, we report the imaging of surface charge dissipation on freshly cleaved mica by Kelvin-probe Force Microscopy (KPFM). A local electrostatic charge micro-domain was generated by friction between an atomic force microscope (AFM) tip and mica, and its decay was tracked by two-dimensional mapping using KPFM. We found time-dependent charge dissipation, which is attributed to the adsorption of water molecules on mica surface.