荧光钙探针试剂及其在生物分析中的应用

本文评论了荧光钙探针试剂合成、性质、结构和生物分析应用方面的进展。各种典型的荧光钙探针试剂均已分别加以讨论。     

荧光试剂Indo-1的合成

以邻硝基酚为原料,经10步反应合成了测定细胞游离钙离子浓度的强荧光试剂--Indo-1,其结构经H NMR,IR和MS表征.     

生物分析中近红外荧光探针进展

本文评述了10余年来方酸菁类、花菁类、噻嗪类、E嗪类、罗丹明类、BODIPY及酞菁等近红外荧光探针的发展。按其种类分别讨论了试剂结构与性能之间的关系,并介绍了它们在生物分析中的某些应用。引用文献42篇。     

罗丹明类荧光染料的合成及应用

本文综述了近年罗丹明类荧光染料的研究进展,详细介绍了它们的结构特征、设计思路、合成方法、目前效果及在生物分析检测领域的应用,并对其未来的发展进行了展望。     

罗丹明类荧光染料的合成及应用

本文综述了近年罗丹明类荧光染料的研究进展,详细介绍了它们的结构特征、设计思路、合成方法、目前效果及在生物分析检测领域的应用,并对其未来的发展进行了展望.     

罗丹明类荧光染料的合成及应用

本文综述了近年罗丹明类荧光染料的研究进展,详细介绍了它们的结构特征、设计思路、合成方法、目前效果及在生物分析检测领域的应用,并对其未来的发展进行了展望。     

中位醛基取代Cy5的光谱性能及作为粘度荧光探针的研究

中位醛基取代的五甲川菁染料(Cy5)与传统Cy5相比其最大吸收和发射波长稍微蓝移, 斯托克斯位移增加, 在水中能达到48 nm|具有低的荧光量子产率, 而且随溶剂极性变化较小. 实验发现这种染料对环境的粘度非常敏感, 随着染料分子所处环境的粘度逐渐增加, 其荧光强度明显增加, 而且荧光强度与溶液粘度的关系满足F?rster-Hoffmann关系式. 该染料具有活细胞膜的穿透性, 能跨膜进入活细胞内, 使得该染料在检测活细胞内微环境粘度变化方面具有潜在的应用价值.     

新荧光试剂4-氨基安替比林芳香席夫碱的合成

新荧光试剂4-氨基安替比林芳香席夫碱的合成     

噻唑橙核酸荧光探针的合成及光谱性质研究

本文合成了一类可用于核酸分子检测的噻唑橙类(thiazole orange,TO)菁染料4a、4b和5,并对染料结构进行了表征。其中染料4a未见文献报道。三种染料在Tris-HCl(pH=7.0)缓冲溶液中的最大吸收光谱值分别在504nm、502nm、507nm处。荧光发射光谱表明:染料在Tris-HCl(pH=7.0)缓冲溶液中无荧光,加入ds-DNA后荧光显著增强,荧光增强与加入ds-DNA的量之间呈现良好的线性关系。改变染料取代基后得到的染料4a对ds-DNA表现出更优异的荧光增强性能。     

Optical modulation and selective recovery of Cy5 fluorescence

Fluorescence modulation offers the opportunity to detect low-concentration fluorophore signals within high background. Applicable from the single-molecule to bulk levels, we demonstrate long-wavelength optical depopulation of dark states that otherwise limit Cy5 fluorescence intensity. By modulated excitation of a long-wavelength Cy5 transient absorption, we dynamically modulate Cy5 emission. The frequency dependence enables specification of the dark-state timescales enabling optical-demodulation-based signal recovery from high background. These dual-laser illumination schemes for high-sensitivity fluorescence-signal recovery easily improve signal-to-noise ratios by well over an order of magnitude, largely by discrimination against background. Previously limited to very specialized dyes, our utilization of long-lived dark states in Cy5 enables selective detection of this very common single-molecule and bulk fluorophore. Although, in principle, the "dark state" can arise from any photoinduced process, we demonstrate that cis-trans photoisomerization, with its unique transient absorption and lifetime enables this sensitivity boosting, long-wavelength modulation to occur in Cy5. Such studies underscore the need for transient absorption studies on common fluorophores to extend the impact of fluorescence modulation for high-sensitivity fluorescence imaging in a much wider array of applications.     

A fluorous tag-bound fluorescence derivatization reagent, F-trap pyrene, for reagent peak-free HPLC analysis of aliphatic amines

We have developed a novel pre-column fluorescence derivatization reagent for amines, F-trap pyrene. This reagent comprises a fluorescent pyrene moiety, an amine-reactive Marshall linker, and a fluorophilic perfluoroalkyl group known as fluorous tag. When the reagent reacts with aliphatic amines and amino acids to give fluorescent derivatives, the fluorous tag in the reagent is eliminated simultaneously. Therefore, excess unreacted reagents in the derivatization reaction solution still have the fluorous tag and could be removed by fluorous solid-phase extraction selectively before high-performance liquid chromatography (HPLC) analysis. By using this reagent, 13 kinds of aliphatic amine (C₂–C₁₆) derivatives can be separated within 40 min by reversed-phase HPLC with gradient elution. In this chromatogram, unreacted reagents peak at around 28 min, greatly decrease after fluorous solid-phase extraction, and do not interfere with the quantification of each amine. The detection limits (S/N = 3) for examined aliphatic amines are 3.6–25 fmol per 20 μL injection. We have also applied this reagent successfully to the amino acid analysis.

Organic solvent-free synthesis of Cy39 iodide

The iodide salt of the pyrylium dye Cy39 (Cy39I, 1) has been suggested as an alternative of its perchlorate salt, which is widely used for biolabeling applications. A new efficient organic solvent-free procedure for synthesis of 1, allowing for a greener approach to the cyanine dyes, is suggested. The unique combination of solubility and thermostability of both the starting materials and the target Cy39 salt made the performance of the synthesis in an aqueous solution as well as reusing of unreacted starting materials possible.     

New derivatizing reagent for analysis of diethylene glycol by HPLC with fluorescence detection

N‐(2‐Phenyl‐indolyl)‐acetic acid (PIAA), a new fluorescent derivatizing reagent, was used for the determination of diethylene glycol (DEG) by high‐performance liquid chromatography with fluorescence detection. DEG was derivatized to ester by using PIAA in the presence of 1‐ethyl‐3‐(3‐dimethylaminopropyl)carbodimide hydrochloride (as dehydrating agent) and 4‐(dimethylamino)pyridine (as base catalyst) in acetonitrile at 60°C for 75 min. The influence of solvent, temperature, catalyst base, concentration of labeling reagent, and couple reagent on the derivatization was investigated. The fluorescence detection was performed with excitation at 340 nm and emission at 377 nm. Baseline separation was obtained on an Ultimate XB‐C18 analytical column with water/acetonitrile gradient elution, good linearity was obtained within 0.5–50 μg/mL with a correlation coefficient of 0.9997. The limit of detection was 0.01 μg/mL (signal‐to‐noise ratio = 3). The method has been successfully applied to determine DEG in toothpaste samples with satisfactory recoveries ranging from 89.0 to 94.9%. The proposed method was shown to be a promising technique for the determination of DEG with high sensitivity.     

PAR-palladium as an analytical reagent for hydrogen cyanide: Application to the detection of thiocyanate and to biological and toxicological analysis

Summary PAR-palladium is proposed as a specific, extremely sensitive and reliable reagent for hydrogen cyanide. The reagent has been used to detect hydrogen cyanide in plants, tissue and toxicological material. A new rapid technique for testing thiocyanate in blood and viscera also has been worked out.

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Zusammenfassung Der PAR-Palladium-Komplex wird als spezifisches, äußerst empfindliches und zuverlässiges Reagens für Blausäure vorgeschlagen. Es wurde zum Nachweis in Pflanzen, Geweben und in toxikologischem Material verwendet. Außerdem wurde ein rasches Verfahren zum Nachweis von Rhodanid in Blut und Organen ausgearbeitet.

     

Joint statistical analysis of multichannel time series from single quantum dot-(Cy5)n constructs

One of the major challenges in single-molecule studies is how to extract reliable information from the inevitably noisy data. Here, we demonstrate the unique capabilities of multichannel joint statistical analysis of multispectral time series using F?ster resonance energy transfer (FRET) in single quantum dot (QD)-organic dye hybrids as a model system. The multispectral photon-by-photon registration allows model-free determination of intensity change points of the donor and acceptor channels independently. The subsequent joint analysis of these change points gives high-confidence assignments of acceptor photobleaching events despite the interference from background noise and from intermittent blinking of the QD donors and acceptors themselves. Finally, the excited-state lifetimes of donors and acceptors are calculated using the joint maximum likelihood estimation (MLE) method on the donor and acceptor decay profiles, guided by a four-state kinetics model.     

Direct observation of delayed fluorescence from a remarkable back-isomerization in Cy5

The direct observations of delayed fluorescence and phosphorescence from the cyanine dye Cy5 are reported. The delayed fluorescence is generated from the S(1) state of trans-Cy5 through a reserve intersystem crossing from the cis-triplet state T(1) to the trans-singlet state S(1) via thermal activation. The lowest cis-triplet state is evidenced to be involved in the formation of the isomer. The back-isomerization from cis-triplet state to trans-singlet state crossing, a remarkably back-isomerization pathway that has not been reported before, plays a significant role in this unusual delayed fluorescence.     

Application of Tsunoda reagent to the convenient synthesis of drug-like pyrazoles

The versatility of the under-utilised (cyanomethylene)tributylphosphorane (CMBP, Tsunoda reagent) was demonstrated on two occasions in a drug discovery context. Firstly, the high reactivity of the phosphorane allowed the alkylation of a weakly acidic pyrazole when standard Mitsunobu conditions were unsuccessful. Secondly, the clean reaction profile generally obtained using CMBP allowed the direct use of the crude mixture in a subsequent Suzuki cross coupling. This reagent has utility when isolation of the Mitsunobu reaction product (e.g. containing a boronate) is not desirable.     

The Vilsmeier reagent: a useful and versatile reagent for the synthesis of 2-azetidinones

(Chloromethylene)dimethylammonium chloride (Vilsmeier reagent), prepared easily from N,N-dimethylformamide and oxalyl chloride or thionyl chloride, works as a versatile acid activator reagent for the direct [2+2] ketene-imine cycloaddition of substituted acetic acid and imines in one-pot synthesis under mild conditions. Monocyclic, spirocyclic and 3-electron-withdrawing group β-lactams were synthesized by this method and optimization of conditions were performed.