人食管癌细胞株PTEN的激光共聚焦扫描显微镜检测

目的对人胚食管上皮永生化细胞株SHEE、SHEEMT、食管癌细胞株EC8712中PTEN表达情况进行定量比较和定位观察.方法采用激光共聚焦扫描显微镜光学切片和荧光探针的双重标记技术对三株细胞中PTEN的表达和分布情况进行检测.结果人食管癌细胞中PTEN主要表达在细胞浆和细胞核,在人胚食管癌上皮永生化细胞株SHEE、SHEEMT主要表达在细胞浆,食管癌细胞EC8712中细胞核表达增多,差异有统计学意义(P<0.01);PTEN在三种细胞株中表达强弱顺序为SHEE>SHEEMT>EC8712,差异有统计学意义(P<0.01).结论PTEN在SHEE、SHEEMT和EC8712分化程度不同的细胞株中均表达,表达和分布位置与分化程度相关.     

共聚焦激光扫描显微镜对约氏疟原虫卵囊、子孢子胞内Ca^2＋的检测方法研究

本文报告建立用共聚焦激光扫描显微镜（CLSM）观察约氏疟原虫卵囊,子孢子胞内游离Ca^2 的分布和实验检测方法,以荧光染料下Fluo-3作胞内Ca^2 指示剂,结果显示3u,4umol/L Fluo-3/Am同时加入1ul/ml25%PluronicF-127在37℃恒温下,分别孵育分离的约氏疟原虫子孢子,卵囊60min即可达到最佳的负载效果,子孢子胞内游离C^2 分布均匀,而卵囊内游离C^2 a分布不均匀,成块状分布,囊壁无效荧光,研究表明应用Fluo-3/AM和PluronicF-127结合CLSM技术是研究疟原虫卵囊,子孢子胞浆内游离Ca^2 的准确,灵敏的方法之一。     

REFLECTIVE PROPERTIES OF THE STIGMA IN MALE GAMETES OF ECTOCARPUS SILICULOSUS (PHAEOPHYCEAE) STUDIED BY CONFOCAL LASER SCANNING MICROSCOPY1

The reflection properties of the stigma in male gametes of Ectocarpus siliculosus (Dillw.) Lyngbye were investigated using confocal laser scanning microscopy in the epireflection contrast mode. The complex reflection pattern obtained after optical xy (horizontal) and xz (vertical) sectioning was consistent with stigma ultrastructure as revealed by serial thin sections. The intensity and pattern of the reflection signal varied with the orientation of the cell/stigma to the incident laser light. Maximal reflection occurred only in approximately normal orientation of the stigma to the light source. Focusing of reflected light from an elongated concave depression of the stigma on the region of the flagellar swelling was observed in xy and xz sections of living and fixed gametes. The results indicate the importance of mechanisms (focusing) other than quarter-wave interference reflection in signal amplification by the eyespot of flagellate algae.     

应用激光扫描共聚焦显微镜FRAP技术研究兔早期胚胎发育中细胞间隙连接介导通讯

本研究应用激光扫描共聚焦显微镜的光漂白恢复技术(FRAP)分析兔早期胚胎卵裂球之间通过间隙连接介导的细胞通讯(GJIC)。研究结果发现,用强激光分别将4-细胞期胚胎、异裂胚胎和8-细胞期胚胎的一个卵裂球荧光光漂白后,经过15分钟的荧光恢复,4-细胞期胚胎的光漂白恢复率为17.8％,异裂胚胎的光漂白恢复率为23.7％,二者之间没有明显的差异;8-细胞期胚胎的光漂白恢复率为78.2％,与前二者之间存在明显的差异。推测兔早期胚胎卵裂球细胞间隙连接建立的时间在8-细胞阶段,胚胎卵裂球间隙连接通讯可能是兔胚胎正常发育的重要条件。     

共聚焦显微技术研究ABA诱导蚕豆气孔保卫细胞H_2O_2的产生(简报)

逆境下,植物细胞内ABA含量急剧增加,同时植物也可通过一些酶代谢反应积累活性氧,如H_2O_2,O_2~-。ABA作为逆境信号对气孔运动的显著调节作用已被诸多实验所证实,但关于其对气孔运动调节的细节还知之甚少。H_2O_2作为氧化信号分子在植物抗病信号转导中已得到广泛研究,但H_2O_2是否介导保卫细胞的气孔运动还缺乏直接的证据。我们已初步发现H_2O_2可参与外源ABA诱     

去卵巢致骨质疏松症大鼠腺垂体FS细胞中Cx43、S-100蛋白表达--激光扫描共聚焦显微镜观察

检测间隙连接蛋白Cx43、神经组织蛋白S-100在去卵巢致骨质疏松症(OVX-OP)大鼠腺垂体滤泡星形细胞(FS细胞)中的表达.实验采用10月龄未孕产SD雌性大鼠40只,随机均分为卵巢切除组(OVX组,n=20)和假性手术对照组(Sham组,n=20).于术后6周末用双能X线骨吸收测量法(DEXA)测量大鼠全身及腰椎4-6(L4-6)骨密度(BMD).取两组大鼠垂体,制成连续切片.应用FITC标记的IgG探针,对腺垂体组织中Cx43和S-100进行间接免疫荧光染色,并用激光扫描共聚焦显微镜(LSCM)定位和定量分析腺垂体FS细胞中Cx43、S-100的表达.结果发现,术后6周末OVX组大鼠全身及L4-6BMD均明显低于Sham组值(P<0.01,P<0.01).Cx43阳性荧光反应主要定位于相邻的FS细胞的胞浆中和/或胞膜上.OVX组Cx43阳性表达荧光强度和表达阳性率均显著低于Sham组(P<0.01).S-100蛋白表达定位于FS细胞的胞浆中,两组间S-100阳性表达荧光强度和表达阳性率无显著差异(P>0.05).本研究提示,SD大鼠OVX术后6周出现骨质疏松变化;OVX大鼠腺垂体FS细胞数量无明显变化、而Cx43表达显著下降,后者的变化可能与大鼠OVX-OP发生相关.     

我国正常人共焦激光扫描视神经乳头地形图分析

目的 运用共焦激光扫描检眼镜(LCSO)测定国人(汉族)视神经乳头(ONH)地形图参数的正常值范围,为临床上青光眼的早期诊断及随访提供依据。方法 用HRT(Heidel-berg Retina Tomograph)技术,检测我国(汉族)74只正常眼ONH地形图参数,经统计学处理,制定各参数的均数(Mean)、标准差(SD)、第5、50、95百分位数(P5、P50、P95)值。并分析ONH参数与种族、性别相关的差异;与年龄、眼别相关的差异作了探讨。结果 国人(汉族)ONH参数的正常值(Meen±SD)为视盘面积(DA)2.34±0.46;视杯面积(CA)0.48±0.28;杯盘面积比(CDR)0.2±0.1;盘沿面积(RA)0.18±0.38;视杯体积(CV)0.1±0.1;盘沿体积(RV)0.5±0.19;平均杯深(MeCD)0.18±0.08;最大杯深(MxCD)0.57±0.21;杯形测量(CSM)-0.25±0.06;曲线高度变化(HVC)0.41±0.12;平均视网膜神经纤维层厚度(mRNFLt)0.27±0.08;视网膜神经纤维层交叉段面积(RNFLcsa)1.47±0.48;标准化参考平面高度(RH)0.29±0.11。各参数P5、P50、P95值分别为DA1、59、2.35、3.05;CA0.11、0.41、0.89;CDR0.06、0.19、0.36;RA1.28、1.82、2.49;CV 0.002、0.06、0.29;RV0.25、0.45、0.84;MeCD 0.05、0.18、0.34;MxCD 0.21、0.59、0.95;CSM-0.35、-0.25、-0.14;HVC 0.23、0.4、0.62;mRNFLt0.14、0.27、0.42;RNF     

共聚焦镜观察凋亡巨噬细胞内pH的变化

用透射电镜观察巨噬细胞的形态学改变,结果显示,地塞米松处理８小时后,大部分巨噬细胞发生凋亡特征变化：胞突缩短、减少,胞膜完整。胞体皱缩,胞质密度增加,其中出现大量空泡。胞核染色质边聚、浓缩。另外用激光扫描共聚焦显微镜（ＡＣＡＳ５７０）和ｐＨ荧光探针ＳＮＡＲＦ┐１／ＡＭ实时检测地塞米松处理巨噬细胞胞浆ｐＨ的动态变化。加入地塞米松,多数巨噬细胞胞浆马上发生快速和短期的碱化。随后,胞浆ｐＨ缓慢降低,胞浆酸化。结果表明,胞浆酸化是细胞凋亡发展的必然过程,胞浆碱化则很可能与细胞凋亡的发生相关,也可能与细胞种类、细胞功能状态相关     

TRACKING POLLEN FLOW OF SOLANUM ROSTRATUM (SOLANACEAE) USING BACKSCATTER SCANNING ELECTRON MICROSCOPY AND X-RAY MICROANALYSIS

A technique using micronized metal powders was developed for both general labeling of pollen and marking of individual pollen grains. After labeling, pollen flow is analyzed by the use of backscatter scanning electron microscopy and X-ray microanalysis. To test the efficiency and efficacy of the technique, we assessed differences in pollen distribution in Solanum rostratum, an enantiostylous species with dimorphic anthers which are putatively feeding and pollinating anthers. Pollen from each set of anthers was labeled using different micronized metal powders. We could not confirm the differentiation of functional anthers in S. rostratum. This technique provides an efficient and convenient method for tracking pollen movement within and between flowers, and anthers within a single blossom can be differentially marked.     

ANALYSIS OF SPATIAL DISTRIBUTIONS OF CELLULAR MOLECULES DURING MECHANICAL STRESSING OF CELL SURFACE RECEPTORS USING CONFOCAL MICROSCOPY

Confocal laser scanning microscopy represents a suitable technique to study the localization of cellular components in three dimension. The authors used this technique to analyse cellular events related to mechanical stimulation of integrin receptors on the cell surface. By performing optical sections the distribution of integrin receptors on the apical surface of an osteoblastic cell was determined. Concerning intracellular compartimentalization of signal transduction events, it was demonstrated that mechanical stimulation of integrins induced their linkage to the cytoskeleton. Cytoskeletally associated proteins like vinculin and talin accumulated in the vicinity of the site where the mechanical stress was applied to integrins on the cell surface. Optical sections revealed that clustering of these proteins proceeded to the base of the cell with gradually decreasing extent. In summary, it was demonstrated that the local distribution of cellular components is an important factor in mechanically induced signal transduction.     

SCANNING ELECTRON MICROSCOPY OF ENDOLITHIC ALGAE AND FUNGI USING A MULTIPURPOSE CASTING-EMBEDDING TECHNIQUE

An embedding-casting method was developed to preserve endolithic algal and fungal structures in situ and to make them optically accessible by removal of carbonate matrix. A slow penetration of Epon-812 resin was applied to fill fine boring tunnels and to embed the organisms within the rock. Fixed, dehydrated, embedded organisms were protected from structural damage during the dissolution of the matrix. Remaining Epon-cast reflects boring patterns and surface details. Casts were studied by a scanning electron-microscope and interpreted by an incident light microscope. After being reembedded in Epon-812, the preparations were sectioned, thin-ground and mounted for study by regular light and electron microscopy.     

七星瓢虫（鞘翅目：瓢虫科）触角的扫描电镜观察

用扫描电子显微镜对两性七星瓢虫的触角进行了观察 ,结果显示 ,鳞状触角分七节 ,分别为雄性 95 0 0 0±81 16 μm而雌性为 972 0 0± 9 5 7μm长。电镜下可观察到七星瓢虫触角有 8种类型的感觉器 (sensillae) ,即 :chaetica(Ch) ,trichoidea(Tr) ,basiconica(Ba) ,campaniformia(Ca) ,ampucellaceous(Am) ,scolopalia(Sc) ,placodea(Pl)和钩状感觉器。Ch在雄性长为 6 2 5 0± 2 89而在雌性为 72 5 0± 5 0 0 μm。Tr在雄性长为 10 75± 1 5 0 μm而在雌性为 14 98± 2 2 5 μm长。Ba在雄雌两性中分别为 5 7± 0 0 2 μm和 0 75± 0 0 6 μm长。Ca为半球状 ,其直径在两性中均在 3 0 μm。Am的直径在雄雌中分别为 1 2和 1 5 μm。Ca为椭圆内陷盘状。Sc在其顶部较阔而在末端尖锐 ,2 5 μm。在雄性触角上观察到一钩状感觉器 ,长 2 1 0 μm ,它表明七星瓢虫在触角上的性别两型性。在雄雌中其感觉器Ch ,Ca ,Am ,Pl的数量上接近。Tr在雄性触角上数量较大而Ba则在雌性触角上数量较多。     

具尾蓝隐藻(CHROOMONASCAUDATAGEITLER)的研究 Ⅰ形态特征──光镜及扫描电镜观察

对具尾蓝隐藻（Chrcomonascaudata）进行了光学显微镜及扫描电镜的观察。扫描电镜观察证明具尾蓝隐藻具有明显的纵沟,细胞顶端具圆形四口,表明该种有口沟存在。从而澄清了文献上对该种有无口沟相互矛盾或模糊不清的描述。     

古雪夫三代虫固着器的扫描电镜观察

古雪夫三代虫 (ＧｙｒｏｄａｃｔｙｌｕｓｇｕｓｓｅｖｉＬｉｎｇ)由林氏首次报道[1 ] 。近年来 ,随着南方鲇的人工养殖密度的不断提高 ,古雪夫三代虫引起的疾病愈来愈严重和普遍 ,主要危害 0 5ｋｇ及其以下的个体。其传播途径 ,疾病早期症状 ,在宿主体表的分布和药物防治已有报道[2 ,3] 。为进一步认识与致病作用密切相关的固着器的形态结构 ,本文进行了该三代虫固着器扫描电镜观察。1　材料和方法患三代虫病的南方鲇 (全长 1 0 2— 2 5 5ｃｍ)取自北碚嘉陵江养殖船内的网箱。用手术刀片轻刮病鱼体表 ,将黏液放于灭菌蒸馏水中 ,在双筒解…