In the laboratory experiment part , human peripheral blood , cultured cells and icr mice were study objects . the changes of mitotic chromosome numbers were measured by human metaphase chromosome counts and statistic analyzed used x2 - test . the changes of meiotic chromosome numbers were measured by mice one - cell zygote chromosome counts and statistic analyzed usedx2 - test . the effects of low dose ionizing radiation on the expression of topoisomerase ii were measured by immunocytochemistry , western blot and rt - pcr 流行病学结果显示长期小剂量辐射接触与染色体不分离呈正相关，为进一步在细胞遗传学和分子生物学方面研究小剂量电离辐射与染色体不分离关系及其机制，本课题第二部分以外周血、培养细胞、 icr小鼠为研究对象，用外周血染色体计数和单细胞受精卵染色体计数的方法研究小剂量辐射和拓扑异构酶复旦大学2000级博士生学位论文11a抑制剂及其二者的协同效应对有丝分裂和减数分裂染色体不分离的影响，用免疫细胞化学染色、 westernblot 、 rt pcr等方法研究了电离辐射引起拓扑异构酶a表达变化。
This understand of stored nitrogen compounds restricted seriously the progress in the investigation of vegetative storage proteins . in the dissertation , we studied more extensively the cytology , biochemical properties and biological roles of vegetative storage proteins in swietenia macrophylla and in hevea brasiliensis by light - and electron microscopy , sds - page , page , immuno - blotting , indirect immunohistochemical localization and colloidal gold labelling and cdna clone techniques 采用光镜和电镜技术、 page 、 sds - page和免疫印迹技术、电泳凝胶过碘酸? schiff试剂染色、间接免疫荧光和电镜免疫细胞化学定位技术以及cdna克隆技术，较深入地研究了大叶桃花心木和巴西橡胶树的营养贮藏蛋白质的细胞学、生物化学性质和生物学功能。
In the second trial , this modified discontinuous percoll gradient centrifugation method was introduced to isolate spermatids from the semen of fifteen male infertile patients . then the effect was identified by wright - giemsa stain , flow cytometry analysis , immunocytochemistry and fluorescence in situ hybridization ( fish ) . similary , the 22 % percoll fraction contained mostly haploid cells [ ( 91 . 85 ? 5 . 18 ) % ] ( p < 0 . 005 ) and the mean density in this fraction was ( 1 . 010 ? 0 . 786 ) x 105 / ml C法，对15例各种类型不育患者的精液细胞进行分离，并利用瑞姬染色法、流式细胞术、免疫细胞化学和荧光原位杂交oisffi等方法，从细胞形态特征、 dna倍体、细胞表面标i己与分化抗原，以及原位杂交信号的数目和位置结合细胞核特有的形态等方面加以鉴定。
Methods : in the first part , we survived and cultured m4g3 hybridoma in vitro . after two weeks of injecting hybridoma into mouse abdominal cavity , we collected abundant ascites . at the same time , 4 strains of breast cancer cells had been growing in vitro , digested and collected for immunocytochemistry and western blotting with m4g3 ascites 方法：第一部分：复苏m4g3杂交瘤细胞，体外培养，小鼠腹腔注射，进行单克隆抗体的大量制备；复苏t47d等四种乳腺癌细胞，体外培养，爬片，应用m4g3进行免疫细胞化学实验；四种细胞收集，应用m4g3进行western - blot染色。
The first part of this study is polyphasic taxonomy analysis of the thirteen strains which have herbicidal activities . the polyphasic taxonomy methods include morphology , cytochemistry , dna g + c mol % , phylogenetic analysis of 16s rdna sequences and physiological and biochemical experiments 本实验的第一部分对具有除草活性的13株放线菌( 40001 40013 )进行了系统的多相分类研究，采用了形态学、细胞化学组分、 dnag + cmol 、 16srdna序列分析及生理生化实验等技术方法。
We detected the transcription and translation change of different thrs genes by semiquantitative reverse transcription - polymerse chain reaction ( rt - pcr ) analysisjelisa respectively . at same time the differentiated cell were identified by immunocytochemistry method . results ( l ) nsc induced by mitogens were nestin - positive and incorporated by brdu 以pdgf 、 t _ 3对nsc进行诱导分化，并分别用rt - pcr半定量法、 elisa法检测nsc在不同诱导因子作用下分化前后thr各亚型在转录与翻译水平上的表达变化，用免疫细胞化学方法鉴定分化后的细胞类型。
Injected group , 0 . 1 % saccharin ( 1 . 5 ~ 2ml / rat , in 5min ) intraoral infused group and cta group . the expression of endogenous leucin - enkephalin ( lek ) in the rat brain was observed and 5 parts of the thalamus including laterodorsal thalamic nucleus ( ld ) , lateral part of mediodorsal thalamic nucleus ( mdl ) , ventroposterolateral thalamic nucleus ( vpl ) , ventroposteromedial thalamic nucleus ( vpm ) and reticular thalamic nucleus ( rt ) were comparatively researched before and after the acquisition of cta applying lek - immunocytochemistry . in behavioral experiment , 18 adult male sd rats were divided into normal cta group ( control ) and 2 naloxone i . p 为探讨cta形成过程中enk的作用，本实验用成年雄性sd大鼠35只，分为空白对照组、生理盐水( 2体重)腹腔注射组、 0 . 15mlicl溶液( 2体重)腹腔注射组、 0 . 1糖精溶液口腔灌流组( 1 . 5 - 2ml只， 5min )和cta建立组，采用免疫细胞化学方法，观察了亮脑啡肽( lek )阳性神经元在大鼠脑内的分布情况，并比较了各组大鼠丘脑外侧背核( ld ) 、丘脑内侧背核外侧部( mdl ) 、丘脑腹后外侧核( vpl ) 、丘脑腹后内侧核( vpm )以及丘脑网状核( rt )等5个脑区内lek表达水平的差异；另外将成年雄性sd大鼠18只，分为正常cta建立组以及在cta建立前或cta建立后阿片受体拮抗剂纳洛酮( 2mg kg体重)腹腔注射组，对内源性阿片样物质对于cta建立和保持的影响进行了行为学研究。
Methods : the effects of different neurotrophic factors on the growth and differentiation of neural stem cells were observed by cells counting and immunofluorescence staining . the levels of rara mrna and rxra mrna in differentiated neural stem cells were assayed by rt - pcr . agarose gel electrophoresis and image analysis 方法应用细胞计数和免疫荧光细胞化学法，研究不同神经营养因子对神经干细胞增殖及分化的影响；应用rt - pcr 、琼脂糖凝胶电泳和紫外分光图象分析法检测神经干细胞分化过程中rar和rxr mrna表达量的改变。结果1
To investigate the potential function of nogo - a involved in neuronal development and differentiation , this study mainly used hippocampal neuron culture model , both in conventional and low - density condition , and immunohistochernical and western blot techniques . nogo - a expression pattern in hippocampal neurons at different stages and its subcellular distribution were explored by using specific antibody raised against nogo - a . pc12 , 3t3 cells were also cultured as controls 为探讨nogo a在神经元中的表达与神经元发育分化的关系，本研究借助了海马神经元常规培养方法和低密度条件下培养方法，应用针对nogo a的特异性抗体，采用免疫荧光细胞化学染色和westernblot方法，观察了nogo a在体外培养的大鼠海马神经元表达分布模式，和不同培养时间nogo a亚细胞分布的变化。
The results of these early research work showed that rna polymerase iii transcription was localized in the nucleoplasm . however , with the development and the application of new technologies since 1990s , the controversy arose on the transcription sites of rna polymerase iii . in recent years , more and more scientists presumed that rna polymerase iii transcription might not occur in the nucleoplasm but in the nucleoli 自上个世纪八十年代初期，人们相继运用细胞化学染色、电镜放射自显影等进行研究的结果表明： rna聚合酶的转录发生在核质中，但随着新的研究技术的发展和应用，人们却发现rna聚合酶的转录可能发生在核仁中，从而对早期的研究结果提出了质疑。
