A QUENCHING FLUORESCENCE IMMUNOASSAY METHOD FOR DETERMINATION OF TRACE ALBUMIN USING UNLABELED TERBIUM CHELATE

A fluoroimmunoassay method using unlabeled Terbium chelate is described.The principle is similar to that of fluoroimmunoassay method using lanthanide chelate as labels.The procedure is simpte because labeling process is unnecessary.The recovery of HSA and albumin in urine is 107% and 95% respectively.The standard deviation is tess than 10%.

Clinical role and importance of fluorescence in situ hybridization method in diagnosis of H pylori infection and determination of clarithromycin resistance in H pylori eradication therapy

H pylori is etiologically associated with gastritis, gas-tric and duodenal ulcers, gastric adenocarcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma. Eradicating H pylori may convert rapidly the outcome of related diseases with the use of more accurate diagnostic molecular tests. Indeed some of the tests cannot give the evidence of current infection; H pylori can be detected by noninvasive and invasive methods, the latter requiring an endoscopy. Eradication failure is a big problem in H pylori infection. Recently, clarithromycin resistance in H pylori strains is increasing and eradicati-on therapy of this bacterium is becoming more difficult. Molecular methods have frequently been applied besides phenotypic methods for susceptibility testing to detect clarithromycin resistance due to mutations in the 2143 and 2144 positions of 23S rRNA gene. Fluorescence in situ hybridization (FISH) method on paraffin embedded tissue is a rapid, accurate and cost-effective method for the detection of H pylori infection and to determine clarithromycin resistance within three hours according to the gold standards as a non-culture method. This method can also be applied to fresh biopsy samples and the isolated colonies from a culture of H pylori, detecting both the culturable bacillary forms and the coccoid forms of H pylori, besides the paraffin embedded tissue secti-ons. This technique is helpful for determining the bac-terial density and the results of treatment where clarith-romycin has been widely used in populations to increase the efficacy of the treatment and to clarify the treatment failure in vitro.

Adaptive hp finite element method for fluorescence molecular tomography with simplified spherical harmonics approximation

Recently,the simplified spherical harmonics equations(SP)model has at tracted much att entionin modeling the light propagation in small tissue ggeometriesat visible and near-infrared wave-leng ths.In this paper,we report an eficient numerical method for fluorescence moleeular tom-ography(FMT)that combines the advantage of SP model and adaptive hp finite elementmethod(hp-FEM).For purposes of comparison,hp-FEM and h-FEM are,respectively applied tothe reconstruction pro cess with diffusion approximation and SPs model.Simulation experiments on a 3D digital mouse atlas and physical experiments on a phantom are designed to evaluate thereconstruction methods in terms of the location and the reconstructed fluorescent yield.Theexperimental results demonstrate that hp-FEM with SPy model,yield more accurate results thanh-FEM with difusion approximation model does.The phantom experiments show the potentialand feasibility of the proposed approach in FMT applications.

A TOTAL CONTENT X—RAY FLUORESCENCE METHOD FOR COPPER PROSPECTING

A new method is proposed to prospect copper deposits with portable XRF analyzer. The method is based on the close relation between Cu and the chalcophile elements or some other elements in the geochemical anomalies of a Cu deposit. Applications of the technique in Northeast China are presented.

Influence enhancement effect of bi-frequency ultrasonic irradiation by TA fluorescence method

Based on a previous research of cavitation effect under bi-frequency ultrasound irradiation, this paper studies bi-frequency irradiations with similar experimental settings. The additional irradiation sources with frequencies of 1.04MHz, 0.8MHz and 1.7MHz are individually combined with the main ultrasonic irradiation source with frequency of 28kHz to form bi-frequency ultrasonic irradiation. The intensity of 28kHz irradiation was fixed at 12.5W/cm^2, while the intensity of the ultrasound at the other three frequencies is varied from1 W/cm^2 to 18 W/cm^2. It turns out that under the influence of the bi-frequency irradiation, the fluorescence intensity is obviously greater than the sum of those at individual frequencies. So the frequency of the additional sonication strikingly influences the fluorescence enhancement effect. For example, the fluorescence enhancement effect of 1.04MHz is stronger than that of 1.7MHz, and the enhancement effect of 0.8MHz is further stronger than that of 1.04MHz. Under the sonic intensity of (7.9)W/cm^2, the fluorescence intensity of 1.04MHz is approximately twice that of 1.7MHz while the fluorescence intensity of 0.8MHz is approximately 1.5 times that of 1.04MHz.

Microstructure and UV Fluorescence of Amorphous I/Au Composite Nanofilms Based on LVDCS Method

The amorphous I/Au composite nanofilms were prepared by low vacuum direct current sputtering(LVDCS) method. The optimized preparation technologies contain growth pressure, time, gaseous environment and annealing conditions. The maximum fluorescence emission(λemmax) of I/Au nanofilms was observed at wavelength of 375 nm, and the intensity of fluorescence emission peak of annealed I/Au films was smaller than that of unannealed one due to fewer amorphous Au nanoparticles, caused by annealing treatment. In the UV-Vis absorption spectra, the intensity of UV-Vis absorption peak of annealed I/Au nanofilms is larger than that of the unannealed one. This work also developed a new way to grow I/Au composite fluorescent thin films.

Study on State of Terbium Ion in Liposome by One order Derivative Fluorescence Quenching Method

The state of Tb3+ is investigated in liposome. When the concentration of PC is below CMC (critical micell concentration), most of Tb3+ is associated with PC, the binding constant is about 3.35×103 L/mol. When the concentration of PC is beyond CMC, most of Tb3+ is dimerized, the dimerization constant is about 3.92×104L/mol. In PC?CH?H2O system, the binding constant of Tb3+?CH complex 2.93×104L/mol is obtained.

A New Fluorescence Method for the Measurement of Intracellular Free Rare Earth Ions： Fura－2 as an Indicator

Fura-2 was first used to monitor intracellular RE(3+). The characterizations of RE3+-Fura-2 interactions in solutions simulating intracellular ionic composition show that La3+ and Y3+ both form 1:1 RE(3+)-fura-2 complexes. The apparent dissociation constants are 1.61×10(-7)M,and 4.04×10(-7)M,respectively. The mouse myeloma(SP2/0) cells exposed to free RE(3+) or free RE(3+) and some small molecules in body were Studied by single cell canon measurement system.The results support the assumption that RE(3+) ions cannot enter the cytoplasm of living cells, and the small molecules used in our experiments couldn't improve the permeability of RE(3+)ions.

New HPLC Method with Experimental Design and Fluorescence Detection for Analytical Study of Antihypertensive Mixture,Amlodipine and Valsartan

New HPLC method was developed for determination of amlodipine and valsartan in their binary mixture as a part of routine control of combined formulations. The method was validated to meet official requirements including selectivity, stability, linearity, precision and accuracy. Chromatography was carried out using a LiChrospher RP-18 column, a mixture containing acetonitrile, phosphate buffer of pH 3.5 and methanol (45:45:10, v/v/v) and new fluorescence detection at 255 nm for excitation and 448 nm for emission. The effect of methanol content, pH of the buffer, flow rate, detection wavelengths and column temperature was estimated in robustness study, according to a plan defined by the Plackett-Burman design. For identification of significant effects, both graphical and statistical methods were used. Ro-bustness for dissolution test was checked estimating the effects of paddle speed, temperature and pH of dissolution medium. The method was proved to complying with all official guidelines. Therefore, it is suitable for determination of amlodipine and valsartan in their binary mixtures for different analytical and pharmaceutical purposes.

Determination of Bisoprolol Fumarate by Fluorescence Quenching Method

[Objectives]To establish a new method for indirect determination of bisoprolol fumarate based on fluorescence quenching technology.[Methods]In ammonia water and ammonium chloride buffer solution at pH=9.2,whenλexcitation=277 nm andλemission=596 nm,with the increase of CCu2+,the fluorescence signal intensity of bisoprolol fumarate weakened,and the difference between the fluorescence intensity of bisoprolol fumarate itself and the fluorescence intensity of the test solution after the quencher Cu2+was added(ΔF)and Cbisoprolol fumarate showed a good linear relationship.[Results]In the range of 15.39-76.93μg/mL,ΔF=146.7 Cbisoprolol fumarate+482.1,r=0.9988,and the detection limit is 0.1391μg/mL.[Conclusions]The fluorescence quenching method has been applied to the determination of actual samples with a recovery rate of 99.9%and an RSD of 2.7%.The results are satisfactory.

A FLUORESCENCE QUENCHING IMNUNOASSAY METHOD FOR DETERMINATION OF TRACE ALBUMIN USING UNLABELED EUROPIUM CHELATE

A fluoroimmunoassay method using unlabeled europium chelate is described.The principle is similar to that of fluoroimmunoassay method using lanthanide chelate as labels.The procedure is simple because labeling process is omitted.The detection limit is about 10^(10) mol/L antigen.The relative standard deviation of immunoassay is less than 10%.The recoveries of human serum albumin and estradiol protein conjugate are 96-105% and 111% respectively.

Determination of Anionic Surfactants in Environmental Water Samples Using Fluorescence Enhancement Method Based on Fe_3O_4@PAA-RB Fluorescent Nanoparticles

[ Objective] The study aims at developing a novel fluorescence enhancement method to determine anionic surfactants. [ Method] Based on Fe3O4 @ PAA-RB fluorescent nanoparticles as fluorescent probes, we have developed a novel fluorescence enhancement method for the determi- nation of an anionic surfactant sodium dodecyl sulfate （SDS） through the gradual optimization of experiment conditions. [ Result] Under the opti- mum conditions, the extent of fluorescence enhancement is directly proportional to SDS concentration varying from 0.5 to 16.0 μmol/L, and the de- tection limit reaches 0.051 μmol/L. The relative standard deviation （RSD） for 4.0 μmol/L SDS is 3.3% （ n =6）. The proposed method has been successfully applied to the determination of SDS in environmental water samples, with recovery of 96.3% -105.5%. E Conclusion] The novel fluo- rescence enhancement method is not only simple and rapid, but also has avoided using tedious solvent-extraction and toxic organic solvents.

Determination of Chlorobenzene by Hydroxypropyl-β- Cyclodextrin Sensitized Fluorescence Quenching Method with Neutral Red as a Fluorescence Probe

The fluorescence quenching of inclusion complex of neutral red （NR） and hydroxypropyl-β-cyclodextrin （HP-β-CD） carried by chlorobenzene was investigated. The fluorescence intensity of NR increased due to the formed inclusion complex of HP-β-CD and NR. But the fluorescence intensity of NR-HP-β-CD diminished when chlorobenzene was added, and there was a linear relationship between the fluorescence quenching value of the system （△IF = IF, NR-HP-β-CD - IF, CB-NB-NR-HP-β-CD） and the concentration of chlorobenzene. Based on this, a novel fluorescence quenching method for the determination of chlorobenzene with NR as a fluorescence probe has been developed. Under the optimal conditions, the linear range of calibration curve for the determination of chlorobenzene was 5.0 × 10^-8 - 8.0 × 10^-6 mol/L and the detection limit was 1.0 × 10^-8 mol/L. It has been applied to determination ofchlorobenzene in synthetic waste water samples with satisfactory results.

Determination of heavy metal ion in landfill leachate with solvent extracting ＆ fluorescence quenching method

Add the masking agent and biscyclohexanoneoxalyldihydraone into the diluted clarificd liquid of the landfill leachate which was treatmented by digestion and centrifugal filtration to complexate all heavy metal ion in the trcatmented liquid, and extracted using CHCl3 many time, then it were demasking and decomposing respectively, and adjusted different pH and formed Me^n＋-PAN coordination compound when these metal ion reaction with PAN. The Fluoresence quenching of Rh6G （λex/λem=543mn/558nm） when the metal ion coordination compound was add into the Rh6G solution step by step, the quenching intensity was directly proportional to the concentration of the metal ion in the certain range. So a new method of fast and simple for determination of trace metal ion in landfill lcachate was established to determine metal ion in sample of different landfill leachate in Three Gorge Water Reservoir, and comparison the classic assay method with satisfactory results.

Soller slits automatic focusing method for multi-element fluorescence detector

In X-ray absorption fine structure(XAFS) experiments,Soller slits are widely used as filter devices in order to improve the signal to noise ratio.Performing high accuracy manual focusing operations is a time-consuming process;therefore,this work introduces an automatic focusing method for Soller slits in multi-element fluorescence detectors.This method establishes a relation model between the fluorescence intensity distribution and the coordinates of the fluorescence excitation point.According to this relation model,the actual coordinates of the fluorescence excitation point can be deduced from the detected fluorescence intensity distribution and used in focusing operations.This method has proven to be feasible in an XAFS experiment at the BL14W1 beamline of the Shanghai Synchrotron Radiation Facility.

嫁接方法与套袋处理对山苍子生长、光合特性及叶绿素荧光特征的影响

【目的】比较嫁接方法和套袋处理对山苍子嫁接幼苗生长、光合特性和叶绿素荧光特征的影响,筛选出最适宜的山苍子嫁接方法。【方法】以1年生山苍子实生苗为砧木、3年生山苍子穗条为接穗,分别采用嵌芽接、劈接和切接3种方法嫁接,每种嫁接方法设置套袋和不套袋2个处理,测定不同组合处理嫁接苗的成活率、生长性状指标、叶片光合特性和叶绿素荧光参数,对各指标进行多重比较,并利用模糊隶属函数法对各组合处理的嫁接苗情况进行综合评价。【结果】3种嫁接方法中,不套袋嵌芽接的嫁接成活率最高(93.75%),劈接苗和切接苗经套袋处理的成活率分别较不套袋处理提高9.03%和26.01%;整体以套袋处理成活率稍高。不同山苍子嫁接苗新枝长、新枝基径和叶片数在嫁接后120~150 d达到峰值,其中新枝长以套袋切接苗最大,新枝基径以套袋嵌芽接苗最大,叶片数以不套袋切接苗最大,分别为26.06 cm,5.69 mm和35.55片。与不套袋处理相比,套袋处理嵌芽接和劈接苗叶片的净光合速率(Pn)、蒸腾速率(T_(r))和气孔导度(G_(s))均有所降低,切接苗叶片的P_(n)、T_(r)、G_(s)和水分利用率(WUE)均有所提高;叶片胞间CO_(2)浓度(Ci)以套袋劈接苗最高,为272.11μmol/mol。与不套袋处理相比,套袋处理嵌芽接苗叶片的实际光化学效率(YⅡ)和光化学荧光淬灭系数(qP)以及切接苗PSⅡ最大光合效率(F_(v)/F_(m))、YⅡ和非光化学荧光淬灭系数(NPQ)均有所提高,而嵌芽接苗和劈接苗叶片的F_(v)/F_(m)、NPQ以及切接苗叶片的qP均有所降低。不同组合处理山苍子嫁接苗综合评分从高到低依次为套袋嵌芽接苗>不套袋劈接苗>套袋切接苗>不套袋嵌芽接苗>不套袋切接苗>套袋劈接苗。【结论】山苍子不同组合处理中,以套袋嵌芽接苗和不套袋劈接苗各项指标的综合表现较优,其最适宜的嫁接方法为套袋嵌芽接,建议在江西地区推广应用。

开放性实验:聚乙烯亚胺修饰碳点的合成、纯化与表征

开放性实验是培养拔尖创新人才的有效手段。我们基于学生兴趣、科研热点和实验室条件,设计了实验方案。实验采用一步水热法,以柠檬酸(CA)为碳源,聚乙烯亚胺(PEI)为修饰剂,合成PEI修饰碳点(PEI-CDs)。首先,将CA、PEI在0.1 mol/L的盐酸中超声至完全溶解,混匀后转移到高压反应釜中,放入烘箱130℃反应2 h,制得PEI-CDs原液;然后将冷却至室温的原液旋转蒸发浓缩至2 mL,并用无水乙醇沉淀、洗涤;最后,将所得沉淀放入真空干燥箱,70℃干燥12 h得到PEI-CDs粉末。利用紫外-可见分光光度计、荧光分光光度计、红外光谱仪和透射电镜对所合成的PEI-CDs进行表征。结果表明,所采用的无水乙醇沉淀纯化方法简单、快速、经济、绿色,制得的PEI-CDs水溶性好,发光性能优异,大小均一,稳定性高。通过该开放性实验,学生不仅熟悉了大型仪器的使用,而且增强了科学研究的兴趣。

硬脑膜修复生物膜的大鼠原位脑植入局部神经毒性评价方法

目的采用大鼠原位脑植入模型评价硬脑膜修复生物膜的局部神经毒性,旨在建立脑部原位植入局部神经毒性评价方法。方法选择硬脑膜修复生物膜为研究对象,以上市同类产品为对照品,经大鼠原位脑植入,获取包含颅骨、硬脑膜、植入材料及其周围大脑组织的标本。采用苏木精-伊红(hematoxylin-eosin,HE)染色、Fluoro-Jade C(FJC)特殊染色与胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)免疫荧光考察植入物周围脑组织的局部组织学反应、神经元降解退行性变与星形胶质细胞增生来综合评价植入物的潜在神经毒性风险。结果植入4周、13周、26周后,与对照品相比,供试品的局部组织学反应均为无刺激反应;FJC染色单位面积阳性细胞数量不存在显著性差异(P>0.05),提示供试品的神经元降解程度与对照组相比不存在显著性差异;GFAP染色单位面积阳性细胞数量不存在显著性差异(P>0.05),提示供试品的星形胶质细胞增生程度与对照组相比不存在显著性差异。结论可以采用HE染色、FJC特殊染色与GFAP免疫荧光染色联合评价硬脑膜修复生物膜经大鼠原位脑植入后的局部神经毒性。

牛冠状病毒TaqMan荧光定量PCR检测方法的建立及应用

为建立牛冠状病毒TaqMan荧光定量PCR检测方法,根据GenBank收录的牛冠状病毒AKS-01株N基因序列(KU886219)保守区设计特异性引物和探针,构建重组质粒并进行反应条件优化、特异性试验、重复性试验以及敏感性试验,建立一种检测BCoV的TaqMan荧光定量PCR方法。结果显示,建立的牛冠状病毒TaqMan荧光定量PCR检测方法特异性、敏感性和重复性均良好。该方法BCoV重组质粒标准品在5.75×10^(7)~5.75×10^(3)copies/μL时与Ct值呈现良好线性关系,该方法对牛轮状病毒、牛传染性鼻气管炎病毒、牛病毒性腹泻病毒、牛副流感病毒3型均无交叉反应,特异性良好;该方法对BCoV重组质粒标准品最低检测限为5.75×10^(1)copies/μL;批内和批间重复性试验结果稳定,变异系数均小于2%。利用所建立的TaqMan荧光定量PCR方法对收集的132份样品进行检测,与常规PCR相比,两者符合率为96.21%,可为BCoV的临床检测和流行病学调查提供技术支持。

72例头癣患儿真菌荧光染色与皮肤镜特征分析

目的研究拟通过真菌荧光染色镜检技术结合皮肤镜分析不同临床类型头癣的特点,为临床提供准确、快速诊断头癣的依据。方法收集72例头癣患儿,使用荧光染色法结合皮肤镜观察头癣病发的主要特征;观察随访患儿治疗后荧光染色法及皮肤镜特殊表征是否同时转阴。结果根据临床特征,72例头癣患儿以白癣为主,占58.33%(42/72);荧光染色法阳性率100%,发外孢子占比最高,为62.50%(45/72);皮肤镜诊断的阳性率100%,皮肤镜检查白癣以断发(90.48%)为主、脓癣以脓疱(100%)为主,以逗号发/螺旋发/条码样发为特殊的表征;随访率为59.72%(43/72)。结论真菌荧光染色法结合皮肤镜特征能对头癣疾病及其临床类型做出快速且准确的诊断。