染色法

The biological features of hBMSCs and cryogentic storaged cells were analyzed.hBMSCs memberane antigens were identified by immunofluorescence staining.The potential osteogenous differentiation of hBMSCs was evaluated by the osteogeneic inducer kit.

倒置光学显微镜下观察细胞形态及生长情况，细胞计数法绘制细胞生长曲线，并分析冻存细胞复苏后的生长特性，免疫荧光染色测定hBMSCs表面标志，成骨诱导试剂盒检测hBMSCs向成骨分化的潜能。

We used the avidin-biotin complex technique and EGFR monoclonal antibody to evaluate the expression of EGFR in 29 cases of cholesteatoma and 34 samples of normal postauricular skin. Of patients with cholesteatoma, 79%(23 cases) had EGFR-positive cells in the basal layer, 66%(19 cases) in the parabasal layer, and 62%(18 cases) in the upper layer of the epithelial tissue.

本研究利用免疫组织化学酵素抗体法（avidin-biotin-peroxidase complex，ABC method）以 EGFR 单株抗体免疫染色，评估29例胆脂瘤与34例耳廓后皮肤两者组织上皮内 EGFR 阳性细胞之分布表现。

1Cubic millimeter tissue sampling was fixed with the mix of 1% glutaraldehyde and 4% paraform in 4℃.

利用兔后腿软组织损伤、海水浸泡模型，分别于损伤后浸泡于海水中2小时，浸泡后分别进行各种低渗盐水冲洗，分别于干预处理后1h、3h、6h、8h、12h，在损伤周围0.5cm范围内的肌肉组织中取材，0.5cm3损伤组织用4%福尔马林固定，HE染色，光镜下观察损伤组织的细胞浸润情况，用免疫组化法检测受伤肌肉组织凋亡相关基因Caspase3表达情况；1mm3损伤组织用1%戊二醛一%多聚甲醛混合固定液(pH7.4 ) 4℃固定，电镜观察其细胞凋亡及细胞膜、线粒体等结构的变化；生化检测Na+，K+-ATPase活性，了解其组间的差别。

Methods DNA was ext racted from paraffin-embedded tissue blocks of 44 angioimmunoblastic lymphaden o pathy patients and analyzed with polymerase chain reaction for IgH and TCRγ gene rearrangement. Immunohistochemistry staining was used to detect p53 protein expression.

方法运用聚合酶链反应技术检测44份AIL石蜡包埋组织标本中IgH及TCR γ基因重排情况，采用免疫组织化学ABC法染色技术检测AIL免疫表型及p53蛋白表达。

After the cell growth curves was recorded, RPE cells of the 3-5th passages were utilized. 2、Three different siRNA (siRNAl,siRNA2,siRNA3) targeting against human cx43 gene and one negative control siRNA were designed and transfected into cultured human RPE cells via liposome reagent. The most effective siRNA can be determined by semi-quantitative reverse transcription PCRRT-PCR. 3、To the most effective siRNA, after transfected into human RPEs with different concentration, the cellular proliferate activities were messured by MTT colorimetry ; the percentages of RPE in different cell circle phase was assayed by FCM; the changes of phenotypical properities were observed with SCM; the protein expression of cx43 was studied through immunocytochemistry stain and Weston blot; the communication intercellular was calculated with FRAP; and the ability of recovery was assessed by using an in vitro wound healing model.4、The total proteins of siRNA1 and RPE were seperated by two-dimensional gel electrophoresis and visualized by silver staining. Proteins with significant expression alterations were selected and their peptide mass fingerprints (PMFs were obtained by matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS).The PMFs were used to search NCBInr database by Auto MS-Fit software.

实验方法：1、培养原代的人RPE细胞，经过细胞角蛋白、S-100和神经胶质原纤维酸性蛋白免疫细胞化学鉴定后，通过AO/PI染色技术确定培养细胞的存活率，描记其生长曲线，第3-5代用于以下细胞实验2、生物合成针对人cx43基因的三条小干扰RNA和一条阴性RNA通过脂质体转染RPE细胞后，通过RT-PCR的方法确定抑制效率最高的干扰片断3、将该片段以不同浓度通过阳离子脂质体转染培养的人RPE细胞后，采用MTT法观察其对细胞的增殖力的作用；通过流式细胞仪观察其对细胞周期的影响；通过扫描电镜观察其对细胞形态的影响；通过免疫细胞化学和Weston blot观察其对cx43蛋白表达的作用；采用激光共聚焦和荧光淬灭恢复技术观察荧光恢复速率平均百分率，评价其对细胞间通讯功能的影响；通过制作RPE细胞损伤模型，观察其对损伤修复能力的作用4、分离纯化转染siRNA的RPE组和正常对照组RPE细胞的全部蛋白质，应用等电聚焦电泳和SDS-PAGE双向电泳技术，银染显示分离出的蛋白质斑点，经凝胶图像分析软件对两个样本进行胶图分析，寻找差异蛋白点。

Osteoblasts were detected by alkaline phosphatese and calcium salt staining. The expression of osteopontin was measured by Western blotting.

成骨细胞鉴定采用组织化学方法行碱性磷酸酶和钙化斑块染色同时应用Western blotting法测定骨桥蛋白表达。

The ultrathin cryosections and ethanolic phospho tungstic acid staining sections of rat atrial cardiocytes were prepared. Quantit ative electron probe Xray microanalysis of Hall continuum method was adopted to measure the alteration of Ca content in ASG at the exact time intervals of 15, 30, 60, 120 min after the adrenaline administration.

冷冻超薄切片技术和磷钨酸-乙醇特殊染色技术制备大鼠心房切片。X射线显微定量分析采用Hall连续谱法，用以测量在肾上腺素处理后15、30、60和120 min时ASG中钙含量的变化。

The cell suspensions were injected into the hippocampus of rat which had received unilateral transection of fimbria-fornix before one week. 5,7,14,30,60,90,150 and 180 days after grafting, the rats were perfused and postfixed, cut section. A series of every second and third sections were used for NOS enzyme histochemistry and Nissl histochemistry.

将大鼠14～16d胚胎的基底前脑移植到单侧穹隆海马伞切断的成年大鼠海马内，动物在移植后存活5、7、14、30、60、90、150和180d分别取脑，经NADPH-d法和尼氏染色观察。

Methods: MTT was used to determine the proliferative effects of drug on tumor cells.

MTT法检测硒化壳聚糖对细胞增殖的影响，AO/EB荧光染色用来观察药物对细胞的凋亡作用。

GFAP and S-100protein present strong positive,while Vimentin protein presents weak positive in the fibrillary and protoplasmic astrocytomas.

采用HE染色和S-P法对276例星形细胞瘤中GFAP、S-100、Vimentin的表达情况进行临床病理分析。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。