A Fast and Indirect Fluorescent Antibody Assay for the Vibrio in Large Yellow Croaker Pseudosciaena crocea (Richardson)

A fast and indirect fluorescent antibody assay for the Vibrio alginolyticus and V. parahaemolyticus infecting the large yellow croaker has been developed. The specific antisera for the two strains of vibrio were prepared with New Zealand rabbit and the antiserum and cross-reactive efficacy was tested by coagulation in tube. It showed that the goat anti-rabbit IgG had been labeled by fluorescence isothiocyanate (FITC). The results showed that positive reactions were 100% for the large yellow croaker Pseudosciaena crocea with typical symptom of vibrio infection, while the positive reaction to the pathogen in healthy yellow croakers reached 40%, but seemed negative for aquaculture water. The results demonstrated that this fast and indirect fluorescent antibody assay can be used not only to test the vibrio pathogen in diseased yellow croaker but also in infected animals with no symptom.

Studies on in situ PCR Detected by the BrdU Antibody Technique

Using the BrdU antibody technique followed by an immuno-chemical staining(BAT),the amplification o f DNA fragments specific to human Y chromosome on cell specimen slides was efficiently detected. Whether direct BrdU incorporation into PCR products or in situ hybridization with PCR products on slides, the amplified targetDNA fragments of specimen were visualized by BAT under the microscope. The availability of BAT and differencesin the sensitivity and efficiency between BAT and dig--if-dUTP labeling in cell in situ PCR were disCussed.

The Double Projecting Neuron of the Cat Spinal Dorsal Horn——A Study with Double Retrograde Fluorescent Tracing Technique

The new double projecting neurons were found in the cat spinal dorsal horn by the double retrograde fluorescent tracing technique.Fast Blue(FB)was injected into unilateral dorsal column nucleus(DCN)of adult cats anesthetized with pentobarbital.Nuclear Yellow(NY)was injected ipsilaterally into the lateral cervical nucleus(LCN)8-9 days later.After an additional 18-30 hrs.

Fluorescent Voltage Imaging Technique for the Measurement of Molluscan Neural Activities

The electrophysiological methods using microelectrodes are not appropriate for the simultaneous measurement of neural activities of many neurons. To overcome the difficulty, the fluorescent imaging technique using voltage sensitive dyes can be a powerful technique. The voltage sensitive dyes, however, generally exhibit a relatively small change in their fluorescence intensities, resulting in a low S/N ratio. Additionally, they often exhibit photobleaching and phototoxity. We have therefore improved the fluorescent voltage imaging technique by using a LED as the light source and an electron multiplying (EM)-CCD camera as the fluorescence detector. In this study, we applied our imaging system for the measurement of two kind of molluscan neural activities;one of which is involved in the olfactory processing of the land slug Limax valentianus and the other is involved in the feeding rhythm of the pond snail Lymnaea stagnalis. The system enabled us to measure the neural activities for a long time with a high speed and a high S/N ratio, and the obtained results showed some new physiological findings.

The Preliminary Report on Rumen Protozoa Grazing Rate on Bacteria with a Fluorescence-Labeled Technique

Studies on the bacterial predation rate by rumen protozoa were carried out under laboratory conditions using a technique of fluorescence-labeled bacteria （FLB）. Four Xuhuai goats were used in this experiment to obtain rumen protozoa and bacteria. Two groups were designed as follows： One group was the whole bacteria which were labeled using fluorescence through removing free bacteria from rumen fluid （WFLB）; the other group was the bacteria which were labeled using fluorescence without removing free bacteria from rumen fluid （FLB）. The result indicated that the bacterial predation rates of rumen protozoa was 398.4 cells/（cell h） for the group WFLB, 230.4 cells/（cell h） for the group FLB, when the corresponding values expressed as bacteria-N, they were 2.15 pg N/（cell h） for the group WFLB, and 1.24 pg N/（cell h） for the group FLB, respectively. Extrapolating the assimilation quantity of nitrogen by ciliates on bacteria of Xuhuai goat, there were 103.2 mg N/（d capita） for the group WFLB, and 59.5 mg N/（d capita） for the group FLB, respectively. It was estimated that protein losses due to microbial recycling were 0.645 g pro/（d capita） for the group WFLB and 0.372 g pro/（d capita） for the group FLB, respectively. In addition, the fluorescence-labeled technique would be a potential assay for the determination of bacterial predation rate by rumen protozoa.

Water Quality Monitoring of the Bezerra River （Cascavel, Brazil） Using SR-TXRF Technique

The present study aims to monitor and assess the water quality of the Bezerra River located in the Western Brazilian Parana state. For the monitoring of river waters, six samplings were established per month during one year. As indicators of the water quality, physico-chemical parameters such as water temperature, pH, turbidity, dissolved oxygen and COD （chemical oxygen demand） were chosen, as well as trace and majority element concentrations. It is noteworthy that the mean annual values of conductivity, turbidity and COD have progressively increased along the river with maximum values after the Cascavel western sewage treatment plant. Only 13 elements were found in the six collection points, but the metallic elements Cr, Mn, Fe, Cu and Zn have shown concentrations above the maximum limits recommended by Brazilian environmental legislation, suggesting the presence of highly polluting anthropogenic sources. Correlation analyses were used to determine the spatio-variability of water quality variables. The six collection sites were grouped into two clusters, with the element composition in the first cluster （sites 1, 2 and 6） being due to strong anthropogenic activities. The study of the Bezerra River water quality could help to develop municipal environmental policies and help with the management of water conservation in the Bezerra River basin.

The Application of Quantitative Fluorescence Techniques in China

The introduction of multi-methods of Geo-logging at wellsite has become the major measurement of oil & gas exploration. From the early stage of manually geo-logging to the modern mudlogging with new techonlogies of MWD, LWD and QFT etc. The new technologies have played very important roles in the exploring of oil & gas. Being one of the newest technology of mudlogging, QFT has been widely used in oilfield for about 3 years. When it is put in operation in some oilfields of China in 1997, its advantages in oil & gas detection at wellsite have been continuously recognized,especially in the detection of shows of light oil and condensed oil. A set of powerful classification standard of resource rock oil bearing grades and the interpretation standards have been summarized by the application of the quantitative fluorescencelogging techniques (QFT) in Basins of China, together with gas-logging data, and other information got from the Geo-logging procedures at wellsite.

Comparison of Multiplex Fluorescent PCR with Serum Type-specific Antibody Detection in Diagnosis of Genital Herpes

Objectives: To compare multiplex fluorescent PCRwith serum type-specific antibody detection in thediagnosis of herpes simplex virus (HSV) infection andto evaluate its significance in the diagnosis of genitalherpes.Methods: We detected HSV infection in 121 speci-mens collected from patients with genital herpesusing both multiplex fluorescent PCR and serum type-specific antibody detection. HSV viral isolation wasused as the standard control.Results: When compared with the viral isolation, thesensitivity and specificity for multiplex fluorescentPCR were 100% and 88.89%, respectively afterdiscrepant analysis. The sensitivity and specificity fortype-specific antibody detection was 77.68 % and77.78 %, respectively. However, the type-specificantibody detected HSV in two asymptomatic patientswhile the multiplex fluorescent PCR couldn’t detectany HSV DNA from those specimens.Conclusions: Multiplex fluorescent PCR is a verysensitive and specific method for detection and typingof HSV in the lesion of genital herpes, it failed todetect HSV DNA from the asymptomatic patients.Serum type-specific antibody detection was a lesssensitive and specific test but could detect the specificantibody from some asymptomatic patients. Thecombination of these two techniques would allow rapid,sensitive and accurate detection and typing of HSVand help clinical diagnosis and epidemiologic survey-ing of genital herpes.

Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics

A fluorescent competitive assay for melamine was first developed utilizing dummy molecularly imprinted polymers（DMIPs） as artificial antibodies. This method is based on the competition between fluorescent substances and the unlabeled analyte for binding sites in synthesized DMIPs and the decreased binding of fluorescent substances to DMIPs due to increased concentrations of melamine in the solutions. DMIPs for melamine were synthesized under a hot water bath in the presence of the initiator azobisisobutyronitrile（AIBN） using 2,4-diamino-6-methyl-1,3,5-triazine（DAMT） as a dummy template, methacrylic acid（MAA） as a functional monomer, and ethylene glycol dimethacrylate（EGDMA） as a crosslinking agent. The adsorption capacity and selectivity of DMIPs for melamine were evaluated by the isothermal adsorption curve and Scatchard analysis. The evaluation results showed that the synthesized DMIPs had specific recognition sites for melamine and the maximum adsorption amount was 1 066.33 μg g^（-1）. Later, 5-（4,6-d ichlorotriazinyl） amino fluorescein（DTAF） with a triazine ring, which s lightly resembles m elamine, w as selected as the fluorescent substance. The fluorescent competitive assay using DMIPs as t he antibody mimics was finally established by selecting and optimizing the reaction solvents, DMIPs amount, DTAF concentration, and incubation time. The optimal detection system showed a linear response w ithin range of 0.05-40 mg L^（-1） and the limit of detection（LOD） was 1.23 μg L^（-1）. It was successfully applied to the detection of melamine in spiked milk samples wi th satisfactory recoveries（71.9 to 86.3%）. According to the comparative analysis, the result of optimized fluorescent competitive assay re vealed excellent agreement with the HPLC-MS/MS result for melamine.

Myelin histology:a key tool in nervous system research

The myelin sheath is a lipoprotein-rich,multilayered structure capable of increasing conduction velocity in central and peripheral myelinated nerve fibers.Due to the complex structure and composition of myelin,various histological techniques have been developed over the centuries to evaluate myelin under normal,pathological or experimental conditions.Today,methods to assess myelin integrity or content are key tools in both clinical diagnosis and neuroscience research.In this review,we provide an updated summary of the composition and structure of the myelin sheath and discuss some histological procedures,from tissue fixation and processing techniques to the most used and practical myelin histological staining methods.Considering the lipoprotein nature of myelin,the main features and technical details of the different available methods that can be used to evaluate the lipid or protein components of myelin are described,as well as the precise ultrastructural techniques.

动物源食品中孕酮激素残留的时间分辨荧光免疫分析法的建立

该研究以孕酮为研究对象,设计并制备了新抗原,通过动物免疫得到特异性兔抗体,基于所得抗原和抗体,进一步优化了抗原抗体浓度、反应缓冲液种类及其pH值等关键参数,最终在国内首次建立了用于动物性食品中性激素孕酮残留快速检测时间分辨荧光免疫分析法(TRFIA)。该研究确定最优实验条件为:抗原0.30μg/mL、铕标抗体0.40μg/mL、pH值7.2的TBST工作缓冲液,并建立标准曲线,方法的线性范围为0.46~6.94μg/L,IC_(50)达1.79μg/L,与睾酮交叉反应率为1.25%,与雌二醇、雌三醇等激素无交叉反应。实际样品的平均添加回收率为82.0%~113.0%,批内批间变异系数<5%,与高效液相色谱法(HPLC)检测结果的相关性良好(r2=0.988)。结果表明该研究所建立的TRFIA检测方法准确可靠、特异性好、灵敏度高,完全可用于动物源食品中孕酮残留的快速检测。

燃料油中硫化氢在线检测系统研制与验证

针对当前燃料油中硫化氢检测技术中存在的样品处理过程复杂、仪器损耗大、烃类干扰严重以及测试周期长等问题,提出了一种顶空气相色谱-荧光检测联用技术。利用顶空气相色谱-荧光检测联用技术实现了燃料油硫化氢的实时在线检测。测得的硫化氢浓度的相对标准偏差为3.21%,且其结果重现性较好。实际样品测试结果表明,顶空气相色谱-荧光检测联用技术与船用燃料油国家标准GB 17411—2015规定使用的英国石油学会标准IP 570/15方法测试结果相近,无显著差异,可以满足日常测试需求。同时,该检测方法针对不同理化性质的燃料油均表现出较高的适用性,有望应用于现场快速检测燃料油质量。

基于P32蛋白的山羊痘病毒抗体荧光微球检测方法的建立

为建立一种山羊痘病毒(GTPV)抗体快速检测方法,试验将携带GTPV结构蛋白P32基因的重组质粒pET28a-P32转化至大肠杆菌BL21(DE3)感受态细胞中,经IPTG诱导后获得了重组蛋白,纯化后经Western blot鉴定表明该蛋白具有良好的特异性和反应原性。以荧光微球为标记材料对P32蛋白进行标记,通过优化反应条件制备了快速检测GTPV抗体的荧光微球免疫层析试纸条,并对其性能进行评价。结果:该试纸条与山羊副流感病毒3型(CPIV3)、小反刍兽疫病毒(PPRV)、羊口疮病毒(ORFV)的阳性血清均无交叉反应;阳性血清稀释至800倍仍能检出阳性,敏感性较高;批内变异系数小于10%,批间变异系数小于15%,稳定性较好;与ELISA检测方法对比的总符合率为92.5%。综上,本试验建立的荧光微球试纸条可用于GTPV抗体的快速检测和流行病学调查。

H7亚型禽流感病毒荧光微球抗体检测方法的建立

为建立一种简便、快速、特异的H7亚型禽流感病毒抗体检测方法,将荧光微球偶联标记鼠抗鸡IgG,并将羊抗鼠IgG和重组H7亚型禽流感病毒HA蛋白分别作为质控线(C线)和检测线(T线),喷涂于硝酸纤维素膜上。采用单一变量分析法对反应体系进行了优化,随后对该方法的敏感性、特异性、重复性及符合率进行了评估。结果显示:本研究建立的H7亚型禽流感病毒荧光微球抗体检测方法比参照国标建立的血凝抑制试验灵敏度高2倍;特异性较好,对H5亚型禽流感病毒、H9亚型禽流感病毒、鸡新城疫病毒、鸡传染性支气管炎病毒、鸡传染性法氏囊病病毒和鸡减蛋综合征病毒等常见禽病病原阳性血清的检测结果均为阴性;重复性试验结果的变异系数为3.13%~8.46%,具有较好的重复性;本研究方法和血凝抑制试验血清抗体检测结果的总体符合率为95.67%,两种方法具有较高的一致性。结果表明,本研究建立的H7亚型禽流感病毒荧光微球抗体检测方法性能稳定、结果可靠,为H7亚型禽流感快速检测提供了新方法。

免疫荧光技术在近视相关生物标志物检测中的应用进展

免疫荧光(IF)技术亦称荧光抗体技术,是通过结合荧光团标记的特定抗原或抗体在紫外线发出荧光来实现的一种示踪技术。IF具有极高的灵敏度和信号放大能力,被广泛应用于细菌病毒和寄生虫的鉴别诊断、部分疾病免疫学机制的研究、器官移植的鉴定、抗原组织定位等方面。近视作为一种屈光不正性疾病,严重影响着人们的视力健康,IF在近视机制及治疗措施的研究中发挥了重要作用。本文就近年来IF在近视相关生物标志物检测中的应用进展进行综述。

先进光学诊断技术在含能材料燃烧测试中的应用进展

依据不同光学原理,从光散射、光学发射与吸收和成像三方面综述了激光诱导荧光(LIF)、相干反斯托克斯拉曼散射(CARS)、粒子成像测速(PIV)、可调谐半导体激光吸收光谱(TDLAS)、激光诱导击穿光谱(LIBS)、辐射法、遥感傅里叶变换红外光谱(RS-FTIR)及纹影法等光学诊断技术测试原理及在含能材料燃烧测试中的应用进展;分析了光学诊断技术在燃烧测试中相比于其他传统接触式诊断方法的优越性和各类光学诊断方法的适用性、测量对象及优缺点;展望了微观燃烧产物、火焰温度、燃烧流场速度和火焰结构等测试技术在含能材料燃烧诊断中的发展前景;指出今后的工作应向多诊断方法相结合,发展多维度测量,从而获取更丰富多维的微观数据信息。附参考文献102篇。

基于PLIF检测的生物质挥发分燃烧多环芳烃生成特性

针对生物质挥发分燃烧过程,采用平面激光诱导荧光技术对火焰中1~2环、3~5环多环芳烃(PAHs)的二维分布进行了研究,分析对比了升温速率(1 000 K/min和1 500 K/min)、O_(2)体积分数(21%~50%)等关键因素对木屑、核桃壳颗粒挥发分燃烧过程中PAHs生成与转化特性的影响.结果表明:提高升温速率,生物质挥发分燃烧过程中1~2环和3~5环PAHs信号强度峰值显著增加,同时升温速率提高促进了1~2环和3~5环PAHs向小分子碳氢化合物转化;1~2环和3~5环PAHs信号强度峰值随O_(2)体积分数提高而增加,这是由于增加的O_(2)体积分数促进了PAHs的释放.但是由于O_(2)体积分数的提高同样增加了OH自由基浓度,导致PAHs消耗加快,使得1~2环和3~5环PAHs信号强度持续时间随O_(2)体积分数提高先增加后减少.木屑和核桃壳颗粒挥发分燃烧过程中生成的3~5环PAHs信号强度峰值和信号强度持续时间较为一致,但木屑颗粒生成的1~2环PAHs信号强度峰值更高,这可能是由于木屑颗粒脂肪烃和芳香环含量更高,同时燃烧过程中生成的OH自由基较多,有利于多环PAHs向1~2环PAHs的转化.

Structural and luminescent properties of (Eu,Tb) PO_4·H_2O nanorods/nanowires prepared by microwave technique

Nanowires/nanorods of europium/terbium orthophosphate monohydrate with Eu^3＋ concentration of 6, 11, and 20 at.% were prepared by microwave synthesis method. The effects of Eu^3＋ doping concentration on structure, morphology and optical properties of nanomaterials were also investigated. The results showed that, for all studied Eu^3＋ doping concentrations, a single-crystalline phase of rhabdophane-type （Eu,Tb）PO4·H2O nanowires/nanorods was obtained by using microwave heating of an aqueous solution of terbium（Ⅲ） nitrate, europium（Ⅲ） nitrate and NH4H2PO4 with pH=2. The length and width of these nanowires/nanorods ranged from 150 to 300 nm and from 10 to 50 nm, respectively. The evidence of energy transfer from Tb^3＋ to Eu^3＋ due to the energy overlap between the donor Tb^3＋ and the acceptor Eu^3＋was observed obviously via a significant enhancement in the luminescent intensity of Eu^3＋. Keywords：

Where Is the Site of the “Oxygen Burst” Located During Light Induction in Dark-Adapted Leaves？ A Study Using Photoacoustic Techniques

: The “oxygen burst” phenomenon that appeared during the light-induction period of intact leaves could be monitored using a photoacoustic technique high time resolution. The relationship between oxygen bursts and dark-adapted time, far-red light pretreatment, photothermal signal, and chlorophyll a (Chl a) fluorescence kinetics were investigated in the present study. Using extraneous inhibitors or cofactors of electron transport, a modified vacuum-infiltration method was undertaken to locate directly the site at which oxygen bursts of intact leaves occurred. We found that the photothermal signal showed little evidence of oscillation during the light-induction period. The oxygen burst was resolved into two components if dark-adapted time lasted longer than 20 min. Methyl viologen (MV) or far-red light could not eliminate the first component, whereas formate-Na (pH 7.0,20 μmol/L) eliminated the first component but had no effect on the second one. Furthermore, the photochemical quenching, the electron transport rate of Chl a fluorescence, and the first component of the oxygen bursts approached lowest values simultaneously. This evidence indicates that the site at which the first component of oxygen bursts occurred was located between photo-system (PS)I and PSII (i.e. the PQ pool). The formate-Na experiment also showed a linkage between the first component and the S state of oxygen evolution at the donor side of PSII. Furthermore, elimination of the second component by far-red light and absorption of the second component by MV indicated that the site at which the second component of oxygen bursts may be located at the acceptor side of PSII.

三维垂体基板类器官的构建和鉴定

背景垂体类器官具有广泛的应用前景,但三维垂体基板类器官相关研究十分有限。目的将人诱导多能干细胞(human induced pluripotent stem cells,hiPSCs)在体外三维环境中诱导为三维垂体基板类器官。方法hiPSCs在体外经过悬浮培养形成球状体,并在一系列细胞因子、激活剂或抑制剂的作用下逐步发育成三维垂体基板类器官。定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)检测hiPSCs或类器官中Nanog同源框(Nanog homeobox,NANOG)、SIX同源框1(SIX homeobox 1,SIX1)、配对样同源域1(paired like homeodomain 1,PITX1)、LIM同源框3(LIM homeobox 3,LHX3)mRNA的表达水平,免疫荧光染色检测三维垂体基板类器官中PITX1、LHX3的表达情况。结果hiPSCs在体外经过悬浮培养形成球状体,随着培养时间的延长,类器官形状也从规则的球形变为不规则形。qPCR和免疫荧光染色结果显示类器官在第30、40天分别成功表达垂体基板标志物PITX1和LHX3,这标志着三维垂体基板类器官的成功构建。结论本研究初步构建了三维垂体基板类器官,为垂体发育研究、药物开发和再生医学研究提供了实验室模型。