Anti-epidermal growth factor receptor (anti-EGFR) antibody conjugated fluorescent nanoparticles probe for breast cancer imaging

Fluorescent nanoparticles (FNs) with unique optical properties may be useful as biosensors in living cancer cell imaging and cancer targeting. In this study, anti-EGFR antibody conjugated fluorescent nanoparticles (FNs) (anti-EGFR antibody conjugated FNs) probe was used to detect breast cancer cells. FNs with excellent character such as non-toxicity and photostability were first synthesized with a simple, cost-effective and environmentally friendly modified Stőber synthesis method, and then successfully modified with anti-EGFR antibody. This kind of fluorescence probe based on the anti-EGFR antibody conjugated FNs has been used to detect breast cancer cells with fluorescence microscopy imaging technology. The experimental results demonstrate that the anti-EGFR antibody conjugated FNs can effectively recognize breast cancer cells and exhibited good sensitivity and exceptional photostability, which would provide a novel way for the diagnosis and curative effect observation of breast cancer cells and offer a new method in detecting EGFR.     

水溶性量子点荧光探针用于胃癌细胞相关抗原CA242的检测

基于量子点荧光探针对胃癌细胞相关抗原CA242进行了检测。首先在水溶液中直接合成性能优良的量子点荧光纳米颗粒,并在其表面成功修饰了羊抗小鼠IgG和聚乙二醇,制得功能化的水溶性量子点荧光探针,并利用探针对胃癌细胞相关抗原CA242进行检测,进一步与传统的基于荧光染料标记的免疫荧光分析方法进行了比较。实验结果表明:该功能化的探针能够有效地识别胃癌细胞相关抗原CA242,并且在光稳定性和灵敏度方面都较传统的基于荧光染料标记的免疫荧光分析方法有明显的改善,从而为CA242的相关检测以及胃癌的诊断与愈后判断提供了新的方法。     

An electrochemical method to detect folate receptor positive tumor cells

This paper reports an electrochemical method to detect folate receptor positive tumor cells by making use of the interaction between folic acid immobilized on gold nanoparticles and its receptor over-expressed on tumor cell membrane. Experimental results have shown that a gold electrode modified with folic acid functionalized gold nanoparticles can clearly denote folate receptor positive tumor cells, such as ovarian tumor cells and human cervical cancer cells. So, electrochemical technique has been introduced for cancer cells detection and a simple method to detect folate receptor positive tumor cells has been developed.     

Anti-Her-2 monoclonal antibody conjugated polymer fluorescent nanoparticles probe for ovarian cancer imaging

Fluorescent nanoparticles (FNPs) with unique optical properties may be useful as biosensors in living cancer cell imaging and cancer targeting. A novel kind of polymer fluorescent nanoparticles (PFNPs) was synthesized and its application for ovarian cancer imaging with fluorescence microscopy imaging technology was presented in this study. The PFNPs were synthesized with precipitation polymerization by using methacrylic acid (MAA) as monomer, trimethylolpropane trimethacrylate (Trim) as cross-linker, azobisisobutyronitrile (AIBN) as radical initiator and butyl rhodamine B (BTRB) as fluorescent dye. And the fluorescent dye was embedded into the three-dimensional network of the polymer when the polymer was produced. With this method the PFNPs can be prepared easily. And then the PFNPs were successfully modified with anti-Her-2 monoclonal antibody. The fluorescence probe based on anti-Her-2 monoclonal antibody conjugated PFNPs has been used to detect ovarian cancer cells with fluorescence microscopy imaging technology. The experimental results demonstrate that the anti-Her-2 monoclonal antibody conjugated PFNPs can effectively recognize ovarian cancer cells and exhibit good sensitivity and exceptional photostability, which would provide a novel way for the diagnosis and curative effect observation of ovarian cancer cells.     

水溶性CdTe量子点荧光探针检测肠癌相关T抗原

直接合成性能优良的水溶性CdTe量子点,然后在其表面成功修饰花生凝集素,经过凝胶柱的分离纯化获得功能化的量子点荧光探针.基于T抗原选择性与花生凝集素(PNA)结合的特性,利用该探针对肠癌中高表达的T抗原进行检测,且与传统的荧光染料标记的免疫荧光分析进行了比较.实验结果表明:该功能化的荧光探针能够有效地识别肠癌的相关T抗原,从而为T抗原的检测以及肠癌的临床诊断与愈后判断提供了一种新的方法.     

Intramolecular Charge Transfer (ICT) Based Two-photon Fluorescent Probe for Bisulfite with Bioimaging Applications

Abstract

A two-photon fluorescent probe has been adopted for bisulfite based on an intramolecular charge transfer (ICT) mechanism. The fluorescence intensity of the probe decreased by 837 fold following treatment with HSO₃^? and the detection limit was 42?nM. This probe showed excellent properties such as high photostability, good two-photon properties, and large Stokes shifts. More importantly, this approach offers a rapid, highly selective, and sensitive method to determine HSO₃^? in buffer solutions and real samples. Furthermore, the probe was successfully used for two-photon fluorescence visualization of trace SO₂ derivatives in biological systems.     

Quantitative Fluorescence Ratio Imaging of Intralysosomal Chloride Ions with Single Excitation/Dual Maximum Emission

Fluorescence ratio imaging is currently being used to quantitatively detect biologically active molecules in biosystems; however, two excitations of most existing fluorescent ratiometric probes account for cumbersome operating conditions for imaging. Thus, a fluorescent ratiometric probe, 6‐methoxyquinolinium–dansyl (MQ‐DS), for Cl^? with single excitation/dual maximum emission has been developed. MQ‐DS can preferably localize into lysosomes and display excellent photostability. Upon excitation at a single wavelength, it responds precisely and instantaneously to changes in Cl^? concentrations, and it can be conveniently utilized to implement real‐time fluorescence ratio imaging to quantitatively track alterations in Cl^? levels inside cells treated under various pH conditions, and also in zebrafish with acute wounds. The successful application of the new probe in bioimaging may greatly facilitate a complete understanding of the physiological functions of Cl^?.     

Glycine-conjugated porphyrin fluorescent probe with iRGD for live cell imaging

A porphyrin modified by glycine has been synthesized and developed as a near-infrared(NIR) fluorescence probe to detect tumor.Porphyrins'longwavelength emission at ~650 nm can efficiently avoid the spectral crosstalk with Spontaneous fluorescence in the visible light region.A disulfide-based cyclic RGD peptide named iRGD c(CRGDKGPDC),a tumor homing peptide,harbors a cryptic C-end Rule (CendR)motif that is responsible for neuropilin-1(NRP-1)binding and for triggering extravasation and tumor penetration of the peptide to improve the imaging sensitivity and therapeutic efficacy.We used N-hydroxy succinimide as an activator to introduce the glycine methyl ester to detect tumor.We got a porphyrin modified by glycine.The affinity between probe and tumor cell entered GLC-82 cells(human glandular lung cancer cell line)can be observed by Confocal Microscope.The toxicity of probe has been identified by MTT Assay.The summary has been gotten that the porphyrins were nontoxic to GLC-82 cells and glycine modified porphyrin has a good affinity with GLC-82 cells under the iRGD function by our experiment.     

Differentiation of Normal and Neoplastic Cells by Synchronous Fluorescence: Rat Liver Epithelial and Rat Hepatoma Cell Models

ABSTRACT

In the present study, conventional and synchronous luminescence (SL) were utilized to investigate spectral differences in normal and neoplastic cells. The synchronous fluorescence (SF) method involves scanning simultaneously both emission and excitation wavelengths while keeping a constant wavelength interval between them. This SF procedure simplifies the emission spectrum and provides for greater selectivity and is used to detect subtle differences in the fluorescence emission of the biochemical species of cells from rat tissues. A difference between the fluorescent spectra of the normal rat liver epithelial (RLE) and hepatoma cell lines were detected using synchronous fluorescence. The potential use of SF as a screening tool for cancer diagnosis is discussed.     

荧光/MRI双模态靶向成像诊疗试剂的制备

在聚乙二醇二胺(NH_2-PEG-NH_2)修饰的石墨烯量子点(GODs)表面以酰胺键偶联二乙基三胺五乙酸(DTPA)分子,之后将Gd~(3+)离子与其进行配合,得到了GODs-Gd(DTPA)复合纳米粒子,然后再通过酰胺键在GODs-Gd(DTPA)的表面修饰叶酸(FA)靶分子,最后进一步将阿霉素(DOX)通过π-π堆垛吸附在造影剂的表面,制备了FA/GODs-Gd(DTPA)/DOX荧光/MRI双模态靶向肺癌细胞成像诊疗试剂,通过透射电子显微镜、紫外可见吸收光谱、荧光光谱和激光共聚焦扫描显微镜等手段表征了其形貌、发光性能和靶向成像性能。MRI、激光共聚焦扫描显微镜和MTT等结果表明,相对于正常的HLF细胞,所制备的FA/GODs-Gd(DTPA)/DOX纳米粒子能够靶向检测FA受体高表达的肺癌H460细胞,并具有明显的抗肿瘤活性。     

Piperazine-based new sensor: selective ratiometric sensing of Fe3+, logic gate construction and cell imaging

Newly designed and synthesised chemosensor 1 selectively recognises Fe³⁺ ions in CHCl₃–MeOH (1:1, v/v) by showing ratiometric change in emission and green colouration of the solution under the exposure of UV light. The ensemble 1·Fe³⁺ selectively detects F^?  ions over other halides and the phenomenon is useful to construct combinatorial logic gate. Furthermore, the probe 1 can be used for in vitro detection of Fe³⁺ in human cervical cancer (HeLa) cells.     

A new selective ‘turn-on’ small fluorescent cationic probe for recognition of RNA in cells

Fluorescent imaging probes have revolutionised cell biology by monitoring cellular objects. However, the lack of fluorescent probes with high selectivity for RNA has been a drawback. Thus, selective RNA binding for fluorescent sensors is essential. Here, we report the selective fluorescence enhancement upon addition of RNA. By exploiting a selective recognition of small tetra-cationic probe 1 for RNA, we also explain the possible binding mode for RNA. As a membrane-permeant fluorescence probe, 1 provides selective imaging of RNA not only in human neuroblastoma tumour SH-SY5Y cell line used for Parkinson's disease but also in the unicellular green alga cells. Further exploitation could open new opportunities in neurotoxin and cancer biology.     

A Rotational BODIPY Nucleotide: An Environment‐Sensitive Fluorescence‐Lifetime Probe for DNA Interactions and Applications in Live‐Cell Microscopy

Fluorescent probes for detecting the physical properties of cellular structures have become valuable tools in life sciences. The fluorescence lifetime of molecular rotors can be used to report on variations in local molecular packing or viscosity. We used a nucleoside linked to a meso‐substituted BODIPY fluorescent molecular rotor ( dC^bdp ) to sense changes in DNA microenvironment both in vitro and in living cells. DNA incorporating dC^bdp can respond to interactions with DNA‐binding proteins and lipids by changes in the fluorescence lifetimes in the range 0.5–2.2 ns. We can directly visualize changes in the local environment of exogenous DNA during transfection of living cells. Relatively long fluorescence lifetimes and extensive contrast for detecting changes in the microenvironment together with good photostability and versatility for DNA synthesis make this probe suitable for analysis of DNA‐associated processes, cellular structures, and also DNA‐based nanomaterials.     

A New Highly Selective and Sensitive Assay for Fluorescence Imaging of .OH in Living Cells: Effectively Avoiding the Interference of Peroxynitrite

A new nonredox fluorescent probe to realize the imaging of hydroxyl radicals (^.OH) in living cells was designed and synthesized. The structure comprised the fluorescent dye boron dipyrromethene (BDP) and a 2,2,6,6‐tetramethyl‐1‐piperidinoxyl (TEMPO) unit. This probe could rapidly respond to ^.OH with a detection limit of 18 pM , and it possessed superior photostability and pH insensitivity. Other reactive oxygen species (ROS) and relevant intracellular components did not interfere. In particular, the important problem of ONOO^? interference was efficiently avoided. An MTT assay proved that the probe was not very cytotoxic. The probe could penetrate into intact cell membranes to selectively detect intracellular ^.OH without causing cellular damage in living mice macrophages, normal human liver cells. and human hepatoma cells. These advantageous characteristics make the fluorescent probe potentially useful as a new candidate to detect ^.OH in broad biosystems.     

Sensitive and simple detection of Escherichia coli strain based on time-resolved fluorescence DNA hybridization assay

A two-probe tandem DNA hybridization assay based on time-resolved fluorescence was employed to detect Escherichia coli strain. The amino modified capture probe was covalently immobilized on the common glass slide surface. The Eu(TTA)₃(5-NH₂-phen) with the characteristics of long lifetime and intense luminescence was labeled with reporter probe. The original extracted DNA samples without the purification and amplification process were directly used in the hybridization assay. The concentration of capture probe, hybridization temperature, hybridization and washing time were optimized. The detection limit is about 1.49 × 10³ CFU mL⁻¹E. coli cells, which is comparable to the value of most microbiology methods. The proposed method has the advantages of easy operation, satisfactory sensitivity and specificity, which can provide a promising technique for monitoring the microorganisms.     

A switch-on near-infrared fluorescence-ready probe for Cu(I): live cell imaging

A reaction-based strategy exploiting metal ion mediated oxidative C–O bond cleavage affords selective ‘switch-on’ near-infrared (NIR) emitting cyanine probe for Cu⁺ in aqueous media and live cells. Near-infrared fluorescence-ready probe TPACy readily reacts with Cu⁺ to release the quinone embedded heptamethine cyanine (Cy-quinone) with extended π-electron conjugation responsible for the switch-on NIR fluorescence in aqueous buffer solution (50 mM HEPES, pH 7.2). This probe can be conveniently used for monitoring Cu⁺ without the interference from pH dependent effects of physiological media. Utility of the probe has been demonstrated by its application in the detection of unbound copper species (Cu⁺) in live cells.     

Parametric signal fitting by gaussian peak adjustment: a new multivariate curve resolution method for non-bilinear voltammetric measurements

A novel two-photon fluorescence probe for Pb²⁺ derived from 4-methyl-2,5-dicyano-4′-amino stilbene as a two-photon fluorophore and bis[2-(2-aminophenylsulfanyls)ethyl]amine as a novel Pb²⁺ ligand was developed. The probe possesses small molecule size, large two-photon absorption cross-section (1020 GM), noncytotoxic effect, long-wavelength emission at 609 nm, large Stokes shift (209 nm), excellent photostability, moderate water-solubility, good cell-permeability, and pH-insensitivity in the biologically relevant pH range. The probe can selectively detect Pb²⁺ ions in live cells and living tissues without interference from other metal ions and the membrane-bound probes, and its quenching constant () is 7.58 × 10⁵ M⁻¹.     

Rational Design and Facile Synthesis of Dual-State Emission Fluorophores: Expanding Functionality for the Sensitive Detection of Nitroaromatic Compounds

Six novel benzimidazole-based D-π-A compounds 4 a – 4 f were concisely synthesized by attaching different donor/acceptor units to the skeleton of 1,3-bis(1H-benzimidazol-2-yl)benzene on its 5-position through an ethynyl link. Due to the twisted conformation and effective conjugation structure, these dual-state emission (DSE) molecules show intense and multifarious photoluminescence, and their fluorescence quantum yields in solution and solid state can be up to 96.16 and 69.82 %, respectively. Especially, for excellent photostability, obvious solvatofluorochromic and extraordinary wide range of solvent compatibility, DSE molecule 4 a is a multifunctional fluorescent probe for the visual detection of nitroaromatic compounds (NACs) with the limit of detection as low as 10⁻⁷ M. The quenching mechanism has been proved as the results of photoinduced electron transfer and fluorescence resonance energy transfer processes. Importantly, probe 4 a can sensitively detect NACs not only in real water samples, but also on 4 a -coated strips and 4 a @PBAT thin films.     

靶向性纳米金探针对宫颈癌细胞的激光扫描共聚焦散射成像

制备了粒径均一的纳米金颗粒, 再对其表面进行叶酸修饰, 制得具有靶向性的纳米金探针. 利用激光扫描共聚焦显微镜(LSCM), 对靶向性纳米金的细胞特异性散射成像进行研究. 实验结果表明, 人宫颈癌细胞(Hela)对纳米金-叶酸的摄取作用强于对纳米金的摄取, 但随着时间的延长, 两者的差别逐渐减小. 表明在适当的时间内纳米金-叶酸探针对宫颈癌细胞具有良好的靶向性.     

The synthesis of highly water-dispersible and targeted CdS quantum dots and it is used for bioimaging by confocal microscopy

Synthesis of a highly dispersed hydrophilic CdS nanocrystals and their use as fluorescence labeling for live cell imaging is reported here. By carefully manipulating the surface of CdS nanocrystals, the dispersions of CdS-MAA-PEI-FA nanocrystals with high photostability is prepared. The receptor-mediated delivery of folic acid conjugated quantum dots into folate-receptor-positive cell lines such as CBRH7919 liver cancer cells was demonstrated by confocal microscopy. In the future, the further modified CdS nanoparticles can be used for the tissue imaging in vivo studies.