Quantitating Fluorescence Intensity from Fluorophore: The Definition of MESF Assignment

The quantitation of fluorescence radiance may at first suggest the need to obtain the number of fluorophore that are responsible for the measured fluorescence radiance. This goal is beset by many difficulties since the fluorescence radiance depends on three parameters 1) the probability of absorbing a photon (molar extinction), 2) the number of fluorophores, and 3) the probability of radiative decay of the excited state (quantum yield). If we use the same fluorophore in the reference solution and the analyte then, to a good approximation, the molar extinction drops out from the comparison of fluorescence radiance and we are left with the comparison of fluorescence yield which is defined as the product of fluorophore concentration and the molecular quantum yield. The equality of fluorescence yields from two solutions leads to the notion of equivalent number of fluorophores in the two solutions that is the basis for assignment of MESF (Molecules of Equivalent Soluble Fluorophore) values. We discuss how MESF values are assigned to labeled microbeads and by extension to labeled antibodies, and how these assignments can lead to the estimate of the number of bound antibodies in flow cytometer measurements.     

基于生物发光法的毒性检测仪设计

为了提高食品、工业环境中毒性物质的检测水平和响应时间,设计了一种基于生物发光法的毒性检测仪。该检测仪以生物发光法作为检测手段,利用毒性物质会抑制荧光素强度的原理,通过检测荧光素的强度来计算毒性物质的毒性。实际试验结果表明,系统能有效的监测到荧光的强弱,可快速准确的计算出待测物质的毒性,且响应快速,响应时间仅为30ms;测量精度相对较高,相对误差小于5%。     

Buckwheat trypsin inhibitor enters Hep G2 cells by clathrin-dependent endocytosis

Recombinant buckwheat trypsin inhibitor (rBTI) was studied to evaluate if it could enter cancer cells and to determine the mechanism. Fluorescein isothiocyanate-labelled buckwheat trypsin inhibitor (FITC-BTI) entered Hep G2 cells in a concentration-dependent manner. FITC-BTI colocalised with labelled transferrin (Tf) in the punctate structure, implying that rBTI enters Hep G2 cells by clathrin-dependent endocytosis. Incubation of Hep G2 cells with different chemical inhibitors abolished diffuse, but not punctate fluorescence, thus indicating that membrane potential plays a critical role in this process. Impairment of clathrin-mediated endocytosis by RNAi with clathrin heavy chain greatly reduced or completely abolished both diffuse and punctate fluorescence, further supporting a theory of a single route of endocytosis. Consistent with our working hypothesis, Hep G2 cells which were arrested in the M phase did not show any vesicular or diffuse FITC-BTI. We conclude from these results that both endocytosis and membrane potential are required for rBTI entry into Hep G2 cells.     

在线高分辨率示踪技术在重庆南山隧道水渗漏的应用研究

为研究南山隧道在岩溶槽谷区施工中产生的上覆岩体渗漏,根据试验区的水文地质条件,选择在地表涂山湖水库作为示踪剂投放点,在南山隧道涌水出口处作为接收点,采用荧光素钠作为示踪剂,进行在线高分辨率示踪试验。试验结果表明:涂山湖水库与南山隧道之间很可能存在三个相互并列的岩溶通道,水力连通时间约为142.0min。在线高分辨率示踪技术具有灵敏度高、取样时间密度小和不易受污染等特点。     

Alkylation of Staurosporine to Derive a Kinase Probe for Fluorescence Applications

The natural product staurosporine is a high‐affinity inhibitor of nearly all mammalian protein kinases. The labelling of staurosporine has proven effective as a means of generating protein kinase research tools. Most tools have been generated by acylation of the 4′‐methylamine of the sugar moiety of staurosporine. Herein we describe the alkylation of this group as a first step to generate a fluorescently labelled staurosporine. Following alkylation, a polyethylene glycol linker was installed, allowing subsequent attachment of fluorescein. We report that this fluorescein–staurosporine conjugate binds to cAMP‐dependent protein kinase in the nanomolar range. Furthermore, its binding can be antagonised with unmodified staurosporine as well as ATP, indicating it targets the ATP binding site in a similar fashion to native staurosporine. This reagent has potential application as a screening tool for protein kinases of interest.     

Dendrimer as nanocarrier for drug delivery

Dendrimers are novel three dimensional, hyperbranched globular nanopolymeric architectures. Attractive features like nanoscopic size, narrow polydispersity index, excellent control over molecular structure, availability of multiple functional groups at the periphery and cavities in the interior distinguish them amongst the available polymers. Applications of dendrimers in a large variety of fields have been explored. Drug delivery scientists are especially enthusiastic about possible utility of dendrimers as drug delivery tool. Terminal functionalities provide a platform for conjugation of the drug and targeting moieties. In addition, these peripheral functional groups can be employed to tailor-make the properties of dendrimers, enhancing their versatility. The present review highlights the contribution of dendrimers in the field of nanotechnology with intent to aid the researchers in exploring dendrimers in the field of drug delivery.     

利用萤光染色跟踪水份在润麦时进入小麦颗粒的路径

本文映示了小麦润麦过程中水分在小麦颗粒中扩散的路径．通过将杜伦麦、软麦在含有萤光试剂的溶液中浸泡12,24,48h之后，萤光切片图像显示出色剂．首先是通过麦胚进入麦粒，再由珠心层传送到麦粒的另一端，同时由糊粉层进入到胚乳．色剂沿背部一侧传送要比腹沟一侧快．皮部吸水速度快，而种皮有抵制水份进入胚乳的作用．从微观观测看出，细胞壁(膜)的三维网状结构形成了水份传送的渠道．色剂在细胞壁里的运动是毛细作用，而在胚乳细胞内的运动则是渗透作用．色剂在细胞壁里的运动要快于在胚乳细胞内的运动．色剂在杜伦麦(硬麦)及软麦里的运动模式是相同的，但是色剂进入软麦的速度要比进入杜伦麦快．     

共焦激光眼底造影及其在眼科的应用

共焦激光技术可以记录眼底视网膜、脉络膜血管状况,并造影成像。本文介绍共焦激光眼底造影设备的组成、工作原理、特点及其在眼科的临床应用。     

Preparation and photophysical behaviors of fluorescent chitosan bearing fluorescein: Potential biomaterial as temperature/pH probes

A novel method for attaching fluorescein (via its epoxy derivative) to water‐soluble chitosan, and the temperature/pH‐sensitive qualities of fluorescence were investigated. 3‐Epoxypropoxy fluorescein (EPF) was firstly synthesized by the reaction between fluorescein and epichlorohydrin, and then water‐soluble chitosan bearing fluorescein (CS‐EPF) was prepared via ring‐opening reaction with EPF. They were characterized by the methods of ¹H NMR, MS, IR, UV–Vis, and luminescence spectra, respectively. The chemiluminescent and photophysical behaviors of EPF and CS‐EPF were studied in detail. The results showed that the fluorescent chitosan could still provide temperature and pH sensitivities similar to that of fluo‐ rescein, achieving better long‐term stability and fast equilibrium response. CS‐EPF had an excellent linear response between relative fluorescence intensity and temperature in the range of 0–60°C and two linear relationships between relative fluorescence intensity and pH in 0.0–4.14 and 8.15–13.20, respectively. This investigation may provide a convenient way to prepare low‐cost and multifunctional macromolecule biomaterial to probe pH and temperature in biological systems. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 104: 3960–3966, 2007     

基于荧光素的荧光聚硫酸酯的制备及表征

采用双酚A（BA）、双酚A硫酰氟和荧光素进行高温溶液缩聚反应,制备了荧光聚硫酸酯（PSE-FL）,并对其进行了结构表征及荧光特性研究。结果表明,制备的PSE-FL的数均分子量为33 413 g/mol,重均分子量为55 043 g/mol,分子量分布指数为1.64,为典型的无定形聚合物,分解温度约为319 ℃,玻璃化转变温度约为128 ℃。固体紫外吸收测试结果表明,PSE-FL在502 nm处有最大吸收波长,荧光发射光谱表明PSE-FL在554 nm处显示出荧光发射波长。