分子印迹荧光传感构建及应用

构建一个高灵敏、高选择性检测痕量分析物的传感器广受科研工作者关注。分子印迹技术由于具有高选择性识别、高容量吸附、快速结合、热稳定性以及低成本等优点,已广泛应用于传感构建领域。以分子印迹聚合物为识别单元,结合荧光传感技术所构建的分子印迹荧光传感器在环境污染物痕量检测方面成为研究重点。本文主要介绍分子印迹聚合物的制备方法,总结分子印迹荧光传感器的构建机理和分子印迹荧光传感器在金属离子、有机小分子以及生物大分子检测方面的应用。重点探讨分子印迹传感器在不同数量的荧光团下检测一种或多种目标分析物的方法,包括单一荧光团检测单一目标物、比率荧光检测单一目标物以及分子印迹荧光传感的多元检测。基于以上分析和总结,提出分子印迹荧光传感器的当前挑战和发展前景。     

A Ratiometric Fluorescence Probe for Selective Detection of ex vivo Methylglyoxal in Diabetic Mice

Accurate monitoring of methylglyoxal (MGO) at cell and living level was crucial to reveal its role in the pathogenesis of diabetes since MGO was closely related to diabetes. Herein, a ratiometric fluorescence strategy was constructed based on the capture probe 2,3-diaminonaphthalene (DAN) for the specific detection of MGO. Compared to the fluorescent probes with a single emission wavelength, the ratiometric mode by monitoring two emissions can effectively avoid the interference from the biological background, and provided additional self-calibration ability, which can realize accurate detection of MGO. The proposed method showed a good linear relationship in the range of 0–75 μm for MGO detection, and the limit of detection was 0.33 μm . DAN responded to MGO with good specificity and was successfully applied for detecting the ex vivo MGO level in plasma of KK−Ay mice as a type II diabetes model. Besides, the prepared DAN test strip can be visualized for rapid semi-quantitative analysis of MGO using the naked eye. Furthermore, human skin fibroblasts and HeLa cells were utilized for exogenous MGO imaging, and ex vivo MGO imaging was performed on tissues of KK−Ay mice. All results indicated that the DAN-based ratiometric fluorescence probe can be used as a potential method to detect the level of MGO, thus enabling indications for the occurrence of diabetes and its complications.     

利用聚(N-异丙基丙烯酰胺-co-丙烯酸)银纳米团簇构建宽pH响应范围的激发比率型荧光探针EI北大核心CSCD

以N-异丙基丙烯酰胺(NIPAM)和丙烯酸(AA)为pH敏感单体,合成聚(N-异丙基丙烯酰胺-co-丙烯酸),并以此作为模板制备了pH敏感的聚(N-异丙基丙烯酰胺-co-丙烯酸)银纳米团簇。结合了时间分辨荧光、透射电镜(TEM)、红外光谱(FT-IR)方法,对团簇的光学性能进行了研究。团簇的荧光对不同的pH具有灵敏的响应。团簇的荧光强度与pH在4.95~11.02的范围内呈线性关系,高pH(11.02)与低pH(4.95)相比,荧光强度降低约62.4%。同时,305 nm与453 nm处荧光激发强度的比值(I;/I;)与pH在5.94~11.02的宽范围内成线性关系,利用该荧光比率可以对pH进行定量检测。     

Highly Sensitive Fluorescent pH Microsensors Based on the Ratiometric Dye Pyranine Immobilized on Silica Microparticles

Pyranine (HPTS) is a remarkably interesting pH-sensitive dye that has been used for plenty of applications. Its high quantum yield and extremely sensitive ratiometric fluorescence against pH change makes it a very favorable for pH-sensing applications and the development of pH nano-/microsensors. However, its strong negative charge and lack of easily modifiable functional groups makes it difficult to use with charged substrates such as silica. This study reports a methodology for noncovalent HPTS immobilization on silica microparticles that considers the retention of pH sensitivity as well as the long-term stability of the pH microsensors. The study emphasizes the importance of surface charge for governing the sensitivity of the immobilized HPTS dye molecules on silica microparticles. The importance of the immobilization methodology, which preserves the sensitivity and stability of the microsensors, is also assessed.     

Assembly of Water-soluble AIE-active Fluorescent Organic Nanoparticles for Ratiometric Detection of Hypochlorite in Living Cells

Hypochlorous acid (HOCl) plays a crucial role in many physiological processes and is widely used as bleach, deodorant and fungicide. In this work, we designed an amphiphilic hydrazone fluorescent molecule THG-1 containing hydrophilic sugar units and hydrophobic tetraphenylethylene unit for ratiometric detection of HOCl with high sensitivity and excellent selectivity based on HOCl-triggered hydrolyzation reaction and aggregation-induced emission (AIE) effect. The detection mechanism was verified by liquid chromatograph mass spectrometry experiments and scanning electron microscope (SEM) tests. Contrast experiments revealed that the numbers of lactose unit and hydrazone linker were essential for assembly of THG-1 and detection of HOCl. In addition, THG-1 was successfully used for imaging of exogenous and endogenous HOCl in living cells.     

Ratiometric Electrochemistry: Improving the Robustness,Reproducibility and Reliability of Biosensors

Electrochemical biosensors are an increasingly attractive option for the development of a novel analyte detection method, especially when integration within a point-of-use device is the overall objective. In this context, accuracy and sensitivity are not compromised when working with opaque samples as the electrical readout signal can be directly read by a device without the need for any signal transduction. However, electrochemical detection can be susceptible to substantial signal drift and increased signal error. This is most apparent when analysing complex mixtures and when using small, single-use, screen-printed electrodes. Over recent years, analytical scientists have taken inspiration from self-referencing ratiometric fluorescence methods to counteract these problems and have begun to develop ratiometric electrochemical protocols to improve sensor accuracy and reliability. This review will provide coverage of key developments in ratiometric electrochemical (bio)sensors, highlighting innovative assay design, and the experiments performed that challenge assay robustness and reliability.     

Convenient Determination of Sulfamethazine in Milk by Novel Ratiometric Fluorescence with Carbon and Quantum Dots with On-site Naked-eye Detection and Low Interferences

Sulfamethazine, one of the most widely applied feed additives, has been shown to cause negative health effects to humans. In the present work, a novel and facile fluorescence visual detection probe was established to determine sulfamethazine in milk samples with naked-eye detection. Considering the good stability, excellent optical properties, and easy synthesis, blue-emission carbon dots were used as the standard signal and red-emission CdTe quantum dots as the responsive signal for the determination of sulfamethazine. The fluorescence intensity of red-emission CdTe quantum dots was gradually quenched with increasing concentration of sulfamethazine, while the blue-emission carbon dots response remained constant. Apparent color variations were observed by naked-eye detection in the concentration range from 9.0 to 54?µmol?·?L^?1. In addition, the presented strategy was shown to be promising to provide a rapid, facile, and sensitive method for the determination of sulfamethazine in milk samples with few interferences.     

Ratiometric pH Sensing in Living Cells Using Carbon Dots

The ability to precisely sense physiological pH changes in the cellular environment is exceedingly difficult. Novel technologies are thus required to address this challenge. Fluorescent nanomaterials can be exploited to this effect because their optical properties can exhibit strong pH dependence. Herein, an intracellular pH-sensing probe is developed via a facile microwave-reaction synthesis method for the preparation of carbon dots (CDs) using glutathione and formamide. The CDs possess unique optical properties allowing for concomitant fluorescence in the blue and red regions of the spectrum. These dots are investigated as pH-sensors using the red fluorescence signatures at 650 and 680 nm. The two fluorescence bands respond differently following pH changes in their environment and could thus be used for ratiometric measurements. Cytotoxicity studies of the CDs in glioblastoma cells show no decrease in cell viability up to 100 μg mL⁻¹ (24 h). Fluorescence imaging reveals that the dots localize in lysosomal compartments. Moreover, they can sense changes in lysosomal pH in response to serum and amino acid starvation, as well as administration of diclofenac and metformin, drugs currently in clinical trials for combination treatments of cancer. These CDs offer a new self-referencing approach for live intracellular pH sensing in 2D- and 3D-cell models.