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1.
The differential reflectance and three-dimensional fluorescence spectra of wet and dry wool following UVB irradiation at 303 nm in the presence and absence of oxygen were measured. Fluorescence originating from tryptophan residues was reduced under all irradiation conditions and a nonfluorescent chromophore with a maximum at 320 nm was produced. These findings suggest that neither singlet oxygen nor triplet states are involved in the formation of the chromophore. Following irradiation of wool at 303 nm in the absence of oxygen, fluorescence with an excitation maximum between 360 and 380 nm and an emission maximum at 440 nm appeared. Fluorescence with an excitation maximum between 410 and 420 nm and an emission maximum at 470 nm from a different species was produced by photolysis in the presence of oxygen. The presence of water accelerates the formation of the latter fluorophore and this observation suggests that singlet oxygen is not involved in its formation.  相似文献   

2.
A characteristic feature of the reactivity of indolequinone derivatives, substituents of which can be removed by one-electron reduction under hypoxic conditions, was applied to the development of a new class of fluorescent probes for disease-relevant hypoxia. A reducing indolequinone parent molecule conjugated with fluorescent coumarin chromophores could suppress efficiently the fluorescence emission of the coumarin moieties by an intramolecular electron-transfer quenching mechanism and a conventional internal-filter effect. Under hypoxic conditions, however, the conjugate, denoted IQ-Cou, underwent a one-electron reduction triggered by X irradiation or the action of a reduction enzyme to release a fluorescent coumarin chromophore, whereupon an intense fluorescence emission with a maximum intensity at 420 nm was observed. The one-electron reduction of IQ-Cou was suppressed by molecular oxygen under aerobic conditions. IQ-Cou also showed intense fluorescence in a hypoxia-selective manner upon incubation with a cell lysate of the human fibrosarcoma cell line HT-1080. The IQ-Cou conjugate has several unique properties that are favorable for a fluorescent probe of hypoxia-specific imaging.  相似文献   

3.
Fluorescent silica nanoparticles (FSNPs) can provide high-intensity and photostable fluorescent signals as a probe for biomedical analysis. In this study, FSNPs hybridized with aggregation-induced emission (AIE) luminogens (namely FSNP-SD) were successfully fabricated by a surfactant-free sol-gel method. The FSNP-SD were spherical, monodisperse and uniform in size, with an average diameter of approximately 100 nm, and emitted strong fluorescence at the peak of 490 nm. The FSNP-SD selectively stained the cytoplasmic regions and were distributed in the cytoplasm. Moreover, they can stay inside cells, enabling the tacking of cells over a long period of time. The intracellular vesicles and multinucleated cells were increase gradually with the rise of FSNP-SD concentration. Both cell viability and survival only lost less than 20% when the cells were exposed to the high concentration of 100 μg/mL FSNP-SD. Additionally, the cell apoptosis and intracellular ROS assay indicated that FSNP-SD had no significant toxic effects at the maximum working concentration of 80 μg/mL. This study demonstrated that the FSNP-SD are promising biocompatible fluorescent probes for living cell imaging.  相似文献   

4.
Newly designed and synthesized diarylethene (DAE) derivatives with aliphatic amine sidearms and one with two pyrenes, revealed excellent photo-switching property of central DAE core in MeOH and water. The only exception was bis-pyrene analogue, its DAE core very readily photochemically closed, but reversible opening completely hampered by aromatic stacking interaction of pyrene(s) with cyclic DAE. In this process, pyrene fluorescence showed to be a reliable monitoring method, an open form characterized by strong emission at 480 nm (typical for pyrene-aggregate), while closed form emitted weakly at 400 nm (typical for pyrene-DAE quenching). Only open DAE-bis-pyrene form interacted measurably with ds-DNA/RNA by flexible insertion in polynucleotide grooves, while self-stacked closed form did not bind to DNA/RNA. For the same steric reasons, flexible open DAE-bis-pyrene form was bound to at least three different binding sites at bovine serum albumin (BSA), while rigid, self-stacked closed form interacted dominantly with only one BSA site. Preliminary screening of antiproliferative activity against human lung carcinoma cell line A549 revealed that all DAE-derivatives are non-toxic. However, bis-pyrene analogue efficiently entered cells and located in the cytoplasm, whereby irradiation by light (315–400 nm) resulted in a strong, photo-induced cytotoxic effect, typical for pyrene-related singlet oxygen species production.  相似文献   

5.
5-aminolevulinic acid (5-ALA)-induced protoporphyrin IX (PpIX) fluorescence is widely used for the intraoperative detection of malignant tumors. However, the fluorescence emission profiles of the accompanying necrotic regions of these tumors have yet to be determined. To address this, we performed fluorescence and high-performance liquid chromatography (HPLC) analyses of necrotic tissues of squamous cancer after 5-ALA administration. In resected human lymph nodes of metastatic squamous cell carcinoma, we found a fluorescence peak at approximately 620 nm in necrotic lesions, which was distinct from the PpIX fluorescence peak at 635 nm for viable cancer lesions. Necrotic lesions obtained from a subcutaneous xenograft model of human B88 oral squamous cancer also emitted the characteristic fluorescence peak at 620 nm after light irradiation: the fluorescence intensity ratio (620 nm/635 nm) increased with the energy of the irradiation light. HPLC analysis revealed a high content ratio of uroporphyrin I (UPI)/total porphyrins in the necrotic cores of murine tumors, indicating that UPI is responsible for the 620 nm peak. UPI accumulation in necrotic tissues after 5-ALA administration was possibly due to the failure of the heme biosynthetic pathway. Taken together, fluorescence imaging of UPI after 5-ALA administration may be applicable for the evaluation of tumor necrosis.  相似文献   

6.
以邻苯二胺和丙二酸为原料,在3 mol/L盐酸催化下合成双(1H-苯并咪唑-2-基)甲烷中间体,然后与月桂醛通过Knoevenagel缩合反应合成了双苯并咪唑衍生物——1,1-双(1H-苯并咪唑-2-基)-1-十三碳烯。利用元素分析、FTIR和~1HNMR对合成产物的结构进行了表征,考察了常规碱催化剂对目标产物收率的影响,并通过UVVis和荧光光谱(FL)测定了目标化合物的光学性能。结果表明:六氢吡啶催化效果最好,产物收率可达76.33%。目标化合物在243、315和361 nm 3处有紫外吸收峰,可有效覆盖日光紫外线UVA和UVB波段;目标化合物溶液在λem=464.5 nm处有蓝紫色荧光,固体薄膜在λ_(em)=476.5 nm处有蓝紫色荧光,荧光量子产率为0.47。标题化合物荧光增白效果与常用荧光增白剂VBL相当。  相似文献   

7.
Near-infrared (NIR)-emitting fluorescent probes are widely used for molecular imaging at the whole-body level. However, NIR-emitting fluorescent probes emitting over λ=700 nm are not suitable for molecular imaging at the cellular level, because most of the conventional fluorescence microscopes have very low optical sensitivity in the NIR region. Thus, to achieve fluorescence imaging at the cellular and whole-body levels by using single probes, visible and NIR-emitting dual-color fluorescent probes are desirable. For dual-color fluorescence molecular imaging, we synthesized fluorescent, recombinant-protein-conjugated, NIR-emitting quantum dots (QDs), in which the recombinant protein consists of enhanced green fluorescent protein (EGFP) and the immunoglobulin binding domain (B1) of protein G. This dual-color fluorescent QD probe binds the Fc region of immunoglobulin G (IgG) through its B1 domain at the QD surface and acts as a molecular-imaging probe at both the cellular and whole-body levels. In this paper, we present the synthesis of fluorescent, recombinant protein (HisEGFP-GB1)-conjugated, NIR-emitting QDs and their application to the dual-color molecular imaging of breast cancer cells in vitro and in vivo.  相似文献   

8.
We report on a new method for the detection of a base at a specific site in a DNA sequence by monitoring the fluorescence emission of fluorescein. To achieve this goal, we developed a new base-discriminating fluorescent (BDF) nucleobase, naphthodeazaadenine ((ND)A). The fluorescence spectrum of the duplex possessing a cytosine base as a complementary base of (ND)A showed a fluorescence peak at 383 nm when using an excitation wavelength of 350 nm. When the complementary base of (ND)A was one of the other bases, the fluorescence intensity was very low. The fluorescence emission spectrum of (ND)A overlapped with the fluorescence excitation spectrum of fluorescein in the wavelength range of 400-500 nm. Thus, we designed FRET-BDF probes containing (ND)A as the FRET donor and fluorescein as the acceptor. The interaction of these two fluorophores, which are separated by defined base pairs, allowed an efficient energy transfer that resulted in a dominant fluorescence emission of fluorescein at 520 nm when using an excitation wavelength of 350 nm. Fluorescence emission from FRET-BDF probes was observed only when the complementary base of (ND)A is C, thus achieving a clear distinction of a C base on the complementary DNA strand. However, the general utility of our method is limited due to the quenching of the (ND)A fluorescence by a G/C base pair flanking (ND)A.  相似文献   

9.
设计合成一种用于pH测定的荧光探针,以罗丹明B、1-0-丙炔基-2,3,4,6-四-0-乙酰基-β-D-葡萄糖苷等为原料,利用Click反应,合成葡萄糖修饰罗丹明B荧光探针,并利用核磁共振波谱对其结构进行表征.进一步研究了该探针的荧光性质,并利用MTT法评价了所制备荧光探针的细胞毒性.实验结果表明,弱酸性条件下,在57...  相似文献   

10.
六羰基钨催化制备聚苯乙炔及其荧光性能的研究   总被引:1,自引:0,他引:1  
用W(CO) 6 催化体系聚合苯乙炔 ,以高产率获得较高相对分子质量的聚苯乙炔。使用UV、IR、GPC等分别对聚苯乙炔结构进行表征。采用不同波长的激发光对聚苯乙炔荧光性能进行了详细研究 ,结果表明 :当聚苯乙炔用 31 0nm激光激发时 ,呈现 3个荧光发光峰 ,峰值分别在348、396和 61 0nm处 ,与其溶液的紫外吸收有类似的光谱曲线特征。荧光峰在 348nm处的荧光 ,当聚苯乙炔质量浓度大于 0 0 5g L时 ,呈现荧光猝灭效应 ;在聚苯乙炔质量浓度为 1 0 - 4g L时 ,呈现出最大的荧光发光量子效率。当激发光波长低于 2 70nm和高于 340nm时 ,348nm波长荧光消失 ;荧光峰在 396nm处的荧光 ,当聚苯乙炔的质量浓度为 1 0 - 2 g L时 ,呈现最大的发光量子效率 ,聚苯乙炔的质量浓度增大或降低 ,其发光强度均呈现减弱现象 ;荧光峰在 61 0nm的荧光 ,在聚苯乙炔质量浓度为 1 0 - 4g L时 ,呈现最大的发光量子效率。荧光性能研究结果证明这种催化剂获得的聚苯乙炔具有多种异构链结构。  相似文献   

11.
Diaminoterephthalates with a maleimide moiety were synthesized and used as fluorescence dyes for sensing thiols. Whereas these "NiWa Blue" dyes showed no emission, the conjugate addition of a thiol to the maleimide group turned on a fluorescence at about 400 nm when irradiating the dye at 338 nm. The neuronal-calcium sensor protein recoverin possesses a single cysteine residue at position 39, which reacts with NiWa Blue, and is therefore labeled by a fluorophore with an emission at about 440 nm. In the absence of Ca(2+), irradiation at 280 nm of a tryptophan residue in close proximity to Cys-bound NiWa Blue lead to strong FRET, which was detected by emission of the dye at 440 nm. In the presence of Ca(2+), the protein holds a conformation with distal Trp and Cys residues, thus FRET of irradiated Trp to Cys-bound NiWa Blue was significantly weakened.  相似文献   

12.
Live-cell sensing of telomerase activity with simple and efficient strategies remains a challenging target. In this work, a strategy for telomerase sensing by using hybridization-sensitive fluorescent oligonucleotide probes is reported. In the presence of telomerase and dNTPs, the designed supporting strand was extended and generated the hairpin structure that catalyzed the next telomerase extending reaction. The special extension mechanism increased the local concentration of another supporting strand and telomerase, which resulted in enhanced telomerase activity. The hybridization-sensitive oligonucleotide probes bound to the hairpin catalyst and generated turn-on fluorescence. This method realized the sensing of telomerase activity in HeLa cell extract with a detection limit below 1.6×10−6 IU μL−1. The real-time in situ observation of telomerase extension was achieved in living HeLa cells. This strategy has been applied to monitor the efficiency of telomerase-targeting anticancer drugs in situ.  相似文献   

13.
以水热法在水相中直接合成了巯基乙酸修饰的CdSe量子点,并将合成的CdSe量子点进行表征、纯化。在波长365nm紫外光的激发下,CdSe量子点发射出明亮的黄绿色荧光,荧光发射峰约位于528nm,将得到的CdSe量子点纳米发光材料应用于非渗透性客体上潜指纹的荧光标记成像研究,发现CdSe量子点溶液显现的手印纹线流畅,显现细节特征明显,呈现明亮的黄绿色荧光指纹,具有很高的实用价值和鉴定价值。  相似文献   

14.
Oligonucleotide hybridization probes that fluoresce upon binding to complementary nucleic acid targets allow the real‐time detection of DNA or RNA in homogeneous solution. The most commonly used probes rely on the distance‐dependent interaction between a fluorophore and another label. Such duallabeled oligonucleotides signal the change of the global conformation that accompanies duplex formation. However, undesired nonspecific binding events and/or probe degradation also lead to changes in the label–label distance and, thus, to ambiguities in fluorescence signaling. Herein, we introduce singly labeled DNA probes, “DNA FIT probes”, that are designed to avoid false‐positive signals. A thiazole orange (TO) intercalator dye serves as an artificial base in the DNA probe. The probes show little background because the attachment mode hinders 1) interactions of the “TO base” in cis with the disordered nucleobases of the single strand, and 2) intercalation of the “TO nucleotide” with double strands in trans. However, formation of the probe–target duplex enforces stacking and increases the fluorescence of the TO base. We explored open‐chain and carbocyclic nucleotides. We show that the incorporation of the TO nucleotides has no effect on the thermal stability of the probe–target complexes. DNA and RNA targets provided up to 12‐fold enhancements of the TO emission upon hybridization of DNA FIT probes. Experiments in cell media demonstrated that false‐positive signaling was prevented when DNA FIT probes were used. Of note, DNA FIT probes tolerate a wide range of hybridization temperature; this enabled their application in quantitative polymerase chain reactions.  相似文献   

15.
Fluorescence‐based oligonucleotide (ON) hybridization probes greatly aid the detection and profiling of RNA sequences in cells. However, certain limitations such as target accessibility and hybridization efficiency in cellular environments hamper their broad application because RNAs can form complex and stable structures. In this context, we have developed a robust hybridization probe suitable for imaging RNA in cells by combining the properties of 1) a new microenvironment‐sensitive fluorescent nucleobase analogue, obtained by attaching the Lucifer chromophore ( 1,8‐naphthalimide) at the 5‐position of uracil, and 2) a peptide nucleic acid (PNA) capable of forming stable hybrids with RNA. The fluorescence of the PNA base analogue labeled with the Lucifer chromophore, when incorporated into PNA oligomers and hybridized to complementary and mismatched ONs, is highly responsive to its neighboring base environment. Notably, the PNA base reports the presence of an adenine repeat in an RNA ON with reasonable enhancement in fluorescence. This feature of the emissive analogue enabled the construction of a poly(T) PNA probe for the efficient visualization of polyadenylated [poly(A)] RNAs in cells—poly(A) being an important motif that plays vital roles in the lifecycle of many types of RNA. Our results demonstrate that such responsive fluorescent nucleobase analogues, when judiciously placed in PNA oligomers, could generate useful hybridization probes to detect nucleic acid sequences in cells and also to image them.  相似文献   

16.
Intracellular pH plays a significant role in many pathological and physiological processes. A series of quinoline-pyrene probes were synthesized in one-step fashion through an oxonium-ion-triggered alkyne carboamination sequence involving C−C, C−O and C−N bond formation for intracellular pH sensing. The quinoline-pyrenes showed significant red shifts at low pH. Fluorescence lifetime decay measurements of the probes showed decreases in lifetime at pH 4. The probes showed excellent selectivity in the presence of various potential interfering agents such as amino acids and cations/anions. Furthermore, the probes were found to show completely reversible emission behaviour in the window between pH 4 and 7. A morpholine-substituted quinoline-pyrene probe efficiently stained lysosomes with high Pearson correlation coefficients (0.86) with Lysotracker Deep Red DND-99 as a reference. A co-localization study of the probe with Lysotracker DND-99 showed selective intracellular targeting and a shift in fluorescence emission due to acidic lysosomal pH.  相似文献   

17.
采用溶胶凝胶燃烧法合成了具有不同形貌和发光性能的纳米CaNb2O6:Bi3+和纳米CaNb2O6:1%Bi3+,1/2K+。通过调整不同添加剂的量,合成了一些有趣的纳米晶形,如球形、花状和棒状。在254nm紫外光激发下,该材料的发射光谱有两部分组成:一部分是弱的发射带,中心约在360nm;另一部分是410~650nm的较强发射区。在315nm紫外光激发下,该材料表现出350~650nm的宽带发射。透射电子显微镜和荧光光谱证实了纳米晶的形貌和尺寸对材料的发光行为有重大影响,并分析了产生这些影响的原因。  相似文献   

18.
Magnetic resonance imaging (MRI) is a powerful imaging modality, widely employed in research and clinical settings. However, MRI images suffer from low signals and a lack of target specificity. We aimed to develop a multimodal imaging probe to detect targeted cells by MRI and fluorescence microscopy. We synthesized a trifunctional imaging probe consisting of a SNAP-tag substrate for irreversible and specific labelling of cells, cyanine dyes for bright fluorescence, and a chelated GdIII molecule for enhancing MRI contrast. Our probes exhibit specific and efficient labelling of genetically defined cells (expressing SNAP-tag at their membrane), bright fluorescence and MRI signal. Our synthetic approach provides a versatile platform for the production of multimodal imaging probes, particularly for light microscopy and MRI.  相似文献   

19.
光致变色化合物螺萘并吡喃的合成及光谱性能   总被引:1,自引:0,他引:1  
以2-萘酚、苯肼和3-甲基-2-丁酮为原料,设计并合成了一种含萘环螺吡喃类光致变色化合物:1,3,3-三甲基吲哚啉螺萘并吡喃,通过1HNMR、IR和元素分析表征了它的分子结构,利用紫外光谱和荧光光谱对目标化合物的吸收光谱和光致发光光谱进行了研究,初步探索了目标产物的光致变色性能。吸收光谱结果表明,目标分子(闭环体)的二氯甲烷溶液(1×10-4mol/L)在可见光区基本无吸收而呈现无色,但经254 nm的紫外光照射后其开环体在480 nm处有较强的吸收而呈现红色(A=1.1),停止紫外光照射溶液恢复为无色,显示了良好的光致变色性能;发光光谱结果显示,目标分子经紫外光激发后其开环体在433 nm处有较强的蓝光发射。  相似文献   

20.
An NIR emitting (λem≈730 nm) cyanine probe ExCy was synthesized in good yields by extending the π-conjugation length (i. e., with furan moiety) to the donor-accepter system. ExCy exhibited a large Stokes’ shift (Δλ≈100 nm) due to strong intramolecular charge transfer (ICT), and high fluorescence quantum yield (Φfl≈0.47 in DCM). Due to its low fluorescence in an aqueous environment (Φfl≈0.007 in H2O), the probe exhibited the potential of achieving a large fluorescence turn-on upon entering a hydrophobic cellular environment. Fluorescence confocal microscopy studies revealed that ExCy was readily excitable with a far-red laser line (i. e., 640 nm) while the corresponding emission was collected in the NIR region. ExCy exhibited excellent selectivity towards live cell mitochondria according to the co-localization studies. The probe also exhibited high photostability, long-term imaging ability and wash-free staining ability, when being applied to live cells. Our studies indicated that the mitochondrial localization of ExCy was dependent on the membrane potential of the mitochondria. ExCy was successfully utilized as a mitochondrial membrane potential dysfunction indicator to visually identify cells with mitochondrial dysfunction via fluorescence confocal microscopy. ExCy was further examined for potential in vivo imaging of zebrafish.  相似文献   

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