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Studies on in situ PCR Detected by the BrdU Antibody Technique
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作者 Zhang Xiyuan Jiang Haibo +2 位作者 Ma Qi Yang Jianqi Li Li ila Liu Ting (College of Life Sciences, Wuhan University, Wuhan 430072, China)Yan Ju (Genetics Lab of Tong n Medical University, Wuhan 430030) 《Wuhan University Journal of Natural Sciences》 CAS 1996年第1期119-124,共6页
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 i... 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. 展开更多
关键词 BrdU antibody technique cell in situ PCR in situ hybridization human Y chromosome polymerasechain reaction (PCR)
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The Double Projecting Neuron of the Cat Spinal Dorsal Horn——A Study with Double Retrograde Fluorescent Tracing Technique
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作者 Jiao Shoushu Liu Yujun +2 位作者 Wang Yuanshen Zhang Guofeng Lu Guowei 《解剖学报》 CAS 1985年第3期243-243,共1页
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 anest... 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. 展开更多
关键词 A Study with Double Retrograde fluorescent Tracing technique The Double Projecting Neuron of the Cat Spinal Dorsal Horn
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A Fast and Indirect Fluorescent Antibody Assay for the Vibrio in Large Yellow Croaker Pseudosciaena crocea (Richardson) 被引量:1
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作者 王军 苏永全 鄢庆枇 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2003年第1期91-96,共6页
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 prepa... 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. 展开更多
关键词 弧菌 大黄鱼 间接荧光抗体 PseudosciaenaCroces VibrioAlginolyticus VPorahaemolyticus
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Fluorescent Voltage Imaging Technique for the Measurement of Molluscan Neural Activities
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作者 Yuuta Hamasaki Makoto Hosoi +2 位作者 Shogo Nakada Tomoya Shimokawa Minoru Saito 《Open Journal of Biophysics》 2013年第1期54-58,共5页
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 vol... 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. 展开更多
关键词 fluorescent VOLTAGE Imaging technique NEURAL Activity MOLLUSK S/N ratio OLFACTORY Processing Feeding RHYTHM
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The Preliminary Report on Rumen Protozoa Grazing Rate on Bacteria with a Fluorescence-Labeled Technique 被引量:7
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作者 WANG Meng-zhi WANG Hong-rong +2 位作者 LI Guo-xiang CAO Heng-chun LU Zhan-jun 《Agricultural Sciences in China》 CAS CSCD 2008年第6期768-774,共7页
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 obta... 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. 展开更多
关键词 rumen protozoa grazing rate bacteria-N fluorescence-labeled technique
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The Application of Quantitative Fluorescence Techniques in China
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作者 Hu Yutao Yang Haibo Guo Qingxia 《Global Geology》 2000年第2期220-222,共3页
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 o... 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. 展开更多
关键词 QUANTITATIVE FLUORESCENCE APPLICATION techniqueS
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Water Quality Monitoring of the Bezerra River (Cascavel, Brazil) Using SR-TXRF Technique
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作者 Phallcha Luízar Obregón Fernando Rodolfo Espinoza-Quinones Aparecido Nivaldo Módenes 《Journal of Chemistry and Chemical Engineering》 2014年第6期587-595,共9页
关键词 水质监测 RF技术 巴西 物理化学参数 金属元素 化学需氧量 污水处理厂 时空变异性
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Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics 被引量:1
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作者 DU Xin-wei ZHANG Yan-xin +8 位作者 SHE Yong-xin LIU Guang-yang ZHAO Feng-nian WANG Jing WANG Shan-shan JIN Fen SHAO Hua JIN Mao-jun ZHENG Lu-fei 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2016年第5期1166-1177,共12页
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... 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. 展开更多
关键词 dummy molecularly imprinted polymers MELAMINE fluorescent competitive assay artificial antibody
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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
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作者 刘波 华彦涛 +3 位作者 马楠楠 赵炫 陈丽楠 袁利鹏 《现代食品科技》 CAS 北大核心 2024年第6期279-284,共6页
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... 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。 展开更多
关键词 动物性食品 孕酮 抗体 时间分辨荧光免疫分析 检测
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Myelin histology:a key tool in nervous system research
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作者 Óscar Darío García-García Víctor Carriel Jesús Chato-Astrain 《Neural Regeneration Research》 SCIE CAS CSCD 2024年第2期277-281,共5页
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... 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. 展开更多
关键词 fluorescence microscopy HISTOLOGY light microscopy lipid histochemistry metallographic techniques myelin histochemistry myelin immunohistochemistry myelin structure&composition myelin ultrastructural evaluation tissue fixation&processing
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燃料油中硫化氢在线检测系统研制与验证
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作者 边晖 谢良波 +3 位作者 刘慧琴 刘淑贞 佟丽丽 刘淑民 《化学分析计量》 CAS 2024年第4期101-105,共5页
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... 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。 展开更多
关键词 顶空气相色谱-荧光检测联用技术 燃料油 硫化氢 装置设计
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基于P32蛋白的山羊痘病毒抗体荧光微球检测方法的建立
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作者 孟卫芹 王金良 +5 位作者 吴信明 唐娜 李通 石竞楠 董帅 沈志强 《畜牧与兽医》 CAS 北大核心 2024年第4期89-94,共6页
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... 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。 展开更多
关键词 山羊痘病毒 P32蛋白 荧光微球 抗体
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免疫荧光技术在近视相关生物标志物检测中的应用进展
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作者 赵宇辉 毕宏生 +3 位作者 孙华跃 李文慧 田庆梅 卢秀珍 《中国医药导报》 CAS 2024年第8期36-39,共4页
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... 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。 展开更多
关键词 免疫荧光 免疫荧光染色 荧光抗体 近视 眼科学 免疫学
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先进光学诊断技术在含能材料燃烧测试中的应用进展
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作者 王一平 徐司雨 +4 位作者 姚二岗 李恒 张洋 于瑾 赵凤起 《火炸药学报》 EI CAS CSCD 北大核心 2024年第1期1-16,共16页
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... 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。 展开更多
关键词 物理化学 含能材料燃烧 光学诊断技术 激光诱导荧光 激光吸收光谱 纹影法
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Structural and luminescent properties of (Eu,Tb) PO_4·H_2O nanorods/nanowires prepared by microwave technique 被引量:2
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作者 Nguyen Thanh Huong Nguyen Duc Van +4 位作者 Dinh Manh Tien Do Khanh Tung Nguyen Thanh Binh Tran Kim Anh Le Quoc Minh 《Journal of Rare Earths》 SCIE EI CAS CSCD 2011年第12期1170-1173,共4页
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, m... 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: 展开更多
关键词 microwave technique nanowires/nanorods energy transfer fluorescence spectroscopy rare earths
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Where Is the Site of the “Oxygen Burst” Located During Light Induction in Dark-Adapted Leaves? A Study Using Photoacoustic Techniques 被引量:3
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作者 Da-YongFAN Rong-FuGAO 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2005年第5期567-578,共12页
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... 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. 展开更多
关键词 chlorophyll a (Chl a) fluorescence extraneous inhibitors and cofactors of electron transport oxygen burst of intact leaves photoacoustic technique
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三维垂体基板类器官的构建和鉴定
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作者 王胜杰 姜得悦 +2 位作者 秦巧臻 江小霞 郭清华 《解放军医学院学报》 CAS 2024年第6期624-630,共7页
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... 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。 展开更多
关键词 诱导多能干细胞 垂体 类器官 聚合酶链反应 荧光抗体技术
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聚合物胶束体内命运分析技术的研究进展
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作者 于璐瑶 刘美辰 +3 位作者 郭莹霞 尤建嵩 史美云 尹磊 《化学试剂》 CAS 2024年第7期75-84,共10页
聚合物胶束(PMs)是研究非常广泛的纳米给药系统。它是由两亲性聚合物组成的自聚集纳米颗粒,这些聚合物在临界胶束浓度(CMC)以下以单分子形式存在于溶液中,在CMC以上则会自组装形成胶束。相对于其他纳米载体,PMs具有独特的性质:相对较小... 聚合物胶束(PMs)是研究非常广泛的纳米给药系统。它是由两亲性聚合物组成的自聚集纳米颗粒,这些聚合物在临界胶束浓度(CMC)以下以单分子形式存在于溶液中,在CMC以上则会自组装形成胶束。相对于其他纳米载体,PMs具有独特的性质:相对较小的尺寸,疏水性核对疏水性化合物具有良好溶解性,亲水性壳能延长药物在血液的循环时间。对PMs体内时空命运的未知,使得PMs从实验室设计到临床应用的成功转化仍然面临重大挑战。目前,一系列分析测试技术已经开发出来用于揭示PMs的体内命运,包括放射性示踪,荧光共振能量转移(FRET)、聚集诱导发射(AIE)、聚集荧光猝灭(ACQ)、液质联用(LC-MS),透射电子显微镜(TEM)等。主要叙述了一系列分析技术在PMs体内命运研究中的进展。 展开更多
关键词 聚合物胶束 体内命运 荧光生物成像 分析技术 研究进展
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小反刍兽疫病毒非结构蛋白C单克隆抗体的制备与鉴定
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作者 李菊 毕冬琳 +5 位作者 杨晓莉 杨东亮 张潇文 刘方程 李琼毅 柏家林 《浙江农业学报》 CSCD 北大核心 2024年第5期1047-1054,共8页
为制备小反刍兽疫病毒(PPRV)非结构蛋白C单克隆抗体,根据PPRV C基因编码多肽链氨基酸序列抗原性分析,设计合成一条含20个氨基酸的抗原肽(CRSGKPRGETPGPLLPEIMQ)和一条含21个氨基酸的筛选抗原多肽(PLRAGERGLAPQAVQHRTLIK),将它们分别与... 为制备小反刍兽疫病毒(PPRV)非结构蛋白C单克隆抗体,根据PPRV C基因编码多肽链氨基酸序列抗原性分析,设计合成一条含20个氨基酸的抗原肽(CRSGKPRGETPGPLLPEIMQ)和一条含21个氨基酸的筛选抗原多肽(PLRAGERGLAPQAVQHRTLIK),将它们分别与钥孔血蓝蛋白(keyhole limpet hemocyanin, KLH)和生物素羧基蛋白(biotin carboxyl carrier protein, Biotin)交联,制备获得免疫原和筛选抗原。用免疫原肌肉注射5只8~12周龄、体重约20 g的无特定病原体(specific pathogen free, SPF)级BALB/c雌性小鼠,第1次免疫后分别间隔7 d进行第2次免疫和第3次免疫,三免后21 d进行冲击免疫,冲击免疫后第3天采集血液分离血清,采用间接酶联免疫吸附法(ELISA)测得1只免疫小鼠血清抗体效价为1∶312 500,2只为1∶62 500。取3只小鼠脾细胞与骨髓瘤细胞SP2/0经聚乙二醇(PEG)融合制备杂交瘤细胞,通过间接ELISA筛选出26株阳性淋巴杂交瘤细胞,进一步经克隆化培养筛选出46株单克隆细胞株。通过Western blot(WB)筛选获得2株能稳定分泌特异性抗PPRV C蛋白单克隆抗体的杂交瘤细胞株,WB、间接免疫荧光(indirect immunofluorescence assay, IFA)鉴定显示,制备的单克隆抗体具有较好的灵敏性和病毒反应性。研究结果为进一步阐明C蛋白在PPRV生命周期中的作用奠定了基础,也为小反刍兽疫(PPR)诊断提供了有效的诊断试剂。 展开更多
关键词 小反刍兽疫病毒 非结构蛋白C 杂交瘤细胞技术 单克隆抗体
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色谱技术在抗体分离纯化中的应用进展
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作者 刘家玮 唐长伟 +1 位作者 夏一然 白泉 《色谱》 CAS CSCD 北大核心 2024年第6期533-543,共11页
抗体作为一类重要的生物药物,在疾病诊断和治疗等方面拥有广阔的前景。抗体药物的需求逐年上升,生产规模不断扩大,下游抗体纯化已经成为抗体药物生产的瓶颈。色谱技术具有选择性好、分离效率高等优点,在抗体纯化领域占据主导地位。抗体... 抗体作为一类重要的生物药物,在疾病诊断和治疗等方面拥有广阔的前景。抗体药物的需求逐年上升,生产规模不断扩大,下游抗体纯化已经成为抗体药物生产的瓶颈。色谱技术具有选择性好、分离效率高等优点,在抗体纯化领域占据主导地位。抗体分离纯化流程一般包括样品的粗提、粗提物精制和抗体精纯3个步骤,每个步骤都涉及色谱分离技术。目前已有多种色谱技术(如亲和色谱、离子交换色谱、疏水作用色谱、混合模式色谱以及新型的温敏色谱等)应用于抗体的分离纯化。本文介绍了近年来国内外各种色谱技术在抗体分离纯化方面所取得的研究进展,并对抗体色谱分离技术的未来发展趋势进行了展望。 展开更多
关键词 抗体 色谱技术 分离纯化 亲和色谱 混合模式色谱 温敏色谱 综述
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