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Methods: Mesenchymal stem cells were isolated from human term placenta by digestion of collagenase Ⅱ and their unique growth characteristic of attaching to the wall of cell culture flask. The proliferation ability was detected by living cell number counting and propidium iodide staining. Their surface markers were detected by flow cytometry. The cells were induced to osteoblast with dexamethasone, antiscorbutic acid and β-sodium glycerophosphate. And they also were induced to adipocytes with dexamethasone and insulin. After induction, the cells were observed by Von Kossa staining and oil red O staining.

将人足月胎盘组织经胶原酶Ⅱ消化和贴壁培养法获取间充质干细胞,运用活细胞计数和碘化丙吮检测其增殖能力;采用流式细胞术检测其细胞表面标志的表达;用地塞米松、抗坏血酸及β-磷酸甘油诱导其向成骨细胞分化,并用Von Kossa染色进行鉴定;用地塞米松与胰岛素诱导其向脂肪细胞分化,并以油红O染色进行鉴定。

Methods The immobilizing conditions of neutrophil alkaline phosphatase staining were detected by the stationary liquid of 40%, 60% and 80% acetone-citric acid, and at the same time, the staining conditions were detected at 10, 15 and 20 minutes in the matrix liquid prepared respectively with 2-amino-2-methyl-1.3-propylene glycol (PH9.4-9.6) buffer, barbital (PH9.2) buffer and Tris (PH9.2) buffer.

用40%、60%、80%的丙酮-枸缘酸固定液对中性粒细胞碱性磷酸酶染色固定条件进行测定。同时用2-氨基-2-甲基-1.3-丙二醇(pH9.4-9.6)缓冲液、巴比妥(pH9.2)缓冲液、Tris(pH9.2)缓冲液配制的基质液分别在10、15、20分钟的染色条件进行测定。

In addition to glial cells, CNTF immunoreactivity are widely localized at neurons in the olfactory bulb, cortex, hippocampus, cerebellum, diencephalon and brainstem.

结果表明,CNTF免疫反应物质除广泛存在于中枢神经系统的胶质细胞外,亦有神经元的广泛染色,而且分布广泛,在嗅脑、大脑皮层、海马、小脑、间脑、脑干中均有明显的染色。

Hyphae, loaded with CTC under pH 6. 8 condition and then grown in pH 8. 0 medium for a little while to destroy the apical acidification, could gave very diffuse fluorescence images of CTC membrame bound Ca〓 and vermiform fluorescence spots due to mitochondrias under pH 6. 8 no longer were clearly distinquished, which implied that besides mitochondrias, other cellular organelles with Ca〓 also were stained with CTC. Thus mitochondrias are not the only one intracellular Ca〓 storage, endoplasmic reticulums and Golgi bodies in the same zone may be also the Ca〓 storages. But the extreme apical zone under 2μm of the growing hyphal tip was still almost devoid of stain under pH 8. 0.The CTC fluorescence was concentrated in the subapic zone beyond about 2μm from the tip and then gradually became lower behind about 40μm from the tip. These results did not support the hypothesis which suggested that the cell wall vesicles in the extreme apical zone were intracellular Ca〓 storages.

将菌丝在pH6.8条件下负载CTC,再置于pH8.0培养介质中短暂生长一会儿以消散菌丝顶端的酸化区域,则CTC膜结合Ca〓呈现弥散的荧光影像,在pH 6.8培养条件下显示的蠕虫状的线粒体荧光斑点不再能够清晰辨认,说明除了线粒体之外,还有其它含Ca〓细胞器,也被CTC染色,线粒体并不是细胞内唯一Ca〓库,还可能包括内质网、高尔基体等细胞器,但菌丝最顶端2μm以前的细胞壁泡囊区域不能被染色,最大荧光强度仍位于菌丝顶端2μm以后区域,约45μm以后荧光变弱,实验结果不支持细胞壁泡囊为菌丝胞内Ca〓库的假设。

Results In the heated bone ,bone lacunae were empty ,no bone cells existed ,bone lamella was seen ,granular basophilia staining was seen in some parts of the lamella. In bone medulla , no cell existed , thrombosis occurred in some blood ,partial bone lamella disappeared.

结果 整个视野内骨陷窝空虚,无骨细胞存在,骨板尚清晰,部分骨板出现颗粒状嗜碱性染色,骨髓腔内,基本无细胞结构,血管闭塞,哈佛氏系统结构不清,部分区域骨板层状结构消失,呈一片均质的嗜伊红染色。

Methods: A 5ml bone marrow was extracted from the lilac of human volunteers. By Percoll fluid and density gradient centrifugation, the MSC was obtained; after the cells filled the bottom of vessel, subcultured them, when they subculture in third generation, redigested them, 500 R/min centrifugate, alter the completed medium to chemical definition medium, examined the form change and prolifration of cells by invert microscope, toluidine blue stain、immunocytochemical stain and RT-PCR to test the type Ⅱ collagen mRNA and proteoglycan.

取健康成人髂后上棘处骨髓5ml,经percoll液分离后密度梯度离心,〓/ml密度接种培养,观察原代细胞的贴壁、增殖状况,细胞长满瓶底后进行传代培养;传至第三代细胞,重新消化后以500转/分钟轻度离心5分钟,〓/ml接种,改用化学限定培养基代替完全培养基培养,倒置显微镜观察细胞生长情况及形态变化,甲苯胺蓝染色观察诱导细胞合成细胞外基质中的蛋白多糖,免疫细胞化学染色检测ECM中Ⅱ胶原的蛋白合成,RT-PCR鉴定诱导细胞Ⅱ胶原mRNA的表达。

The aim are:lTo examine the proliferation ability and potential chondroblast differentiation ;2To find an ideal condition stimulated BMSC differentiate into chondroblast;3To examine the chondroblast proliferation in porous scaffolds and to explore the interaction between cells and materials;4To examine the release of cytokine in vitro.

取健康成人略后上棘处骨髓 5ml,经 percoll液分离后密度梯度离心,10'砌l密度接种培养,观察原代细胞的贴壁、增殖状况,细胞长满瓶底后进行传代培养;传至第三代细胞,重新消化后以500转/分钟轻度离心 5分钟,10V加 l接种,改用化学限定培养基代替完全培养基培养,倒置显微镜观察细胞生长情况及形态变化,甲苯胺蓝染色观察诱导细胞合成细胞外基质QCM)中的蛋白多糖,免疫细胞化学染色检测ECM中*胶原的蛋白合成,RT干CR鉴定诱导细胞*胶原mRNA的表达。

Results:17 rats were confirmed successful transplantation by MRI,the capacity of the tumor was 88.40±7.62mm3 on 8th day, 986.80±114.46mm3 on 16th day,the volume growth was 8-12 folds;on 8th day following transplantation,the tumor tissue was gray in color with fish homogen state without interior necrosis, the tumor cells were nest-distributed by HE staining,large dense staining of nucleus with manifest heteromorphism,tumor microvessel count increased by immunohistochemistry with buffy yellow staining, VEGF high expressed in the tumor cells with buffy yellow granule shape.

结果:17只大鼠经MRI检查证实均种植成功,肿瘤体积第8天为88.40±7.62mm3,第16天为986.80±114.46mm3,体积增长8-12倍之间;种植第8天,肿瘤组织呈灰白色、鱼肉均质状,内部未见坏死,HE染色肿瘤细胞呈巢状分布,胞核大浓染,异型性明显;免疫组织化学示肿瘤微血管数目较多,呈棕黄色染色,肿瘤细胞VEGF高表达,呈棕黄色细颗粒状。

By means of experiments of marker transfer,acridine orange sensitivity test,F' curring and mini-chromosome transformation it is.conchi-ded that the F' plasmid is always in an integrated state in the Sin strains and that the initiation of the replication of the bacterial chromosome is carrie...

标记转移、吖啶橙敏感性,F′质粒消除和mini-染色体质粒转化等实验说明,Sin菌株中F′质粒始终处于整合状态,并且在40℃中细菌染色体的复制由整合状态的F′质粒所带动。

By means of experim ents of marker transfer,acridine orange sensitivity test,F' curring and mini-chr omosome transformation it is concluded that the F' plasmid is always in an integ rated state in the SIn strains and that the initiation of the replication of the bacterial chromosome is carried on by the integrated F' plasmid.

标记转移、吖啶橙敏感性、F′质粒消除和mini-染色体质粒转化等实验说明,Sin菌株中F′质粒始终处于整合状态,并且在40℃中细菌染色体的复制由整合状态的F′质粒所带动。

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