一种新的在线细胞显微观察仪及其在生化反应器上的应用研究

针对生化反应器应用条件,提出了用于生化反应器的在线细胞观察仪的基本技术要求。在比较了国内外现有的细胞在线显微工作原理后,研制了一种基于暗视场的新的显微细胞观察仪,介绍了其关键技术及结构。另外,原位在线显微细胞观察仪应用于酿酒酵母和哺乳动物细胞HEK293细胞的培养试验,在线细胞计数结果与离线细胞计数和细胞干重相比较,均有很好的相关性,表明此仪器基本满足使用要求。     

一新用于生化反应器的在线细胞显微观察仪

针对生化反应器应用条件,提出了用于生化反应器的在线细胞观察仪的基本技术要求。在比较了国内外现有的细胞在线显微工作原理后,研制了一种基于暗视场的新的显微细胞观察仪,介绍了其关键技术及结构。另外,原位在线显微细胞观察仪应用于酿酒酵母和哺乳动物细胞HEK293细胞的培养试验,在线细胞计数结果与离线细胞计数和细胞干重相比较,均有很好的相关性,表明此仪器基本满足使用要求。     

食管癌细胞与正常食管上皮细胞的扫描电镜观察

在透射电镜中,已发现食管癌管细胞与正常食管上皮细胞不仅具有细胞器的差异,而且细胞表面的微绒毛也有不同。由于透射电镜只能观察到细胞的某一切面,而不能观察细胞的立体图像,因之我们应用扫描电镜观察了培养的正常食管上皮与     

利用中性红观察黑藻活细胞的物质进出

植物活细胞可发生质壁分离及质壁分离复原现象,细胞质也处于不断流动的状态(胞质环流)。选用黑藻作为材料,在观察胞质环流现象的同时,观察细胞质壁分离;质壁分离复原实验选用活细胞染色剂中性红(0.1%水溶液)作为指示剂,观察中性红进入细胞的同时,观察复原现象。改进后的实验中可明显观察到中性红进入细胞,从而更好地展示只有"活细胞"才能控制"物质进出"的特点。     

M期细胞选择性电镜取材与观察

从细胞形态学的角度观察,整个细胞的增殖周期唯有分裂期(M 期)细胞的变化显著,便于从形态上区别与鉴定,因此 M 期细胞的变化是细胞形态学研究中极受重视的观察目标。然而,由于其在整个细胞周期中历时最短,在细胞群体中往往只占百分之几的比例,所以要想大量观察 M 期细胞的变化,按过去常规取材的方法是不可能的,尤其是电镜水平的观察,     

介绍一种观察凋亡细胞形态学特征的染色方法--苏木素-伊红染色法(HE染色法)

就苏木素-伊红染色法用于观察与鉴别细胞凋亡与细胞坏死作了较详细的介绍。此染色法对凋亡细胞的形态学特征检测,既方便又可靠,在一般普通光学显微镜下就可进行观察与检测。此法不失于研究细胞凋亡的一种检测观察凋亡细胞形态学特征的简便方法。     

人肝癌细胞与肝星形细胞裸鼠皮下共培养体系的建立

目的:建立裸鼠皮下共培养人肝癌细胞与肝星形细胞模型,观察人肝癌细胞与肝星形细胞间相互作用后超微结构的改变.方法:将16只裸鼠分为两组,肝癌细胞单独培养组和癌细胞与肝星形细胞共培养组,40天后将荷瘤组织切片行光镜及透射电镜观察.结果:肝癌细胞单独培养组中可观察到肝癌细胞的胞质液化及早期细胞凋亡现象,而共培养组中可见肝星形细胞时肝癌细胞的趋化现象,可观察到肝癌细胞结构完整且有增殖趋势.结论:裸鼠皮下荷瘤三维立体模型建立成功,该模型能够模拟肝癌微环境中肝癌细胞与肝星形细胞问的作用,为进一步研究肝癌细胞与肝星形细胞间的相互作用奠定了基础.     

观察洋葱表皮细胞立体形状的简易方法

在植物学教学中,为了使学生能获得关于细胞的感性认识,通常设置观察洋葱表皮细胞基本结构的实验,但由于制作临时切片时通常用水做为介质,所以对细胞的立体形状观察得不十分明显。为了使学生能获得清晰的细胞立体形状的认识,提高实验课的教学效果。现介绍一种观察洋葱...     

荧光显微技术在酿酒酵母细胞研究中的应用

为了探讨荧光显微技术在酿酒酵母细胞不同研究方面的作用,通过GFP标记目标蛋白的同源重组的方法和免疫荧光技术标记两种蛋白,最后利用荧光显微镜观察酵母细胞中某种蛋白定位及两种蛋白共定位情况;分别用荧光染液DAPI、FM4-64、BODIPY、Filipin、DHE和Annexin V试剂处理酵母细胞之后,利用荧光显微镜观察细胞中的细胞核、液泡、脂滴、麦角固醇、ROS和细胞凋亡的情况。结果显示,荧光显微技术在酵母细胞蛋白定位、细胞器观察及细胞中ROS和细胞凋亡等研究方面具有重要作用。     

一种酶标板转子低速离心细胞制片方法

常规培养悬浮细胞Jurkat,离心收集细胞后,通过载玻片直接涂片、细胞涂片离心机和酶标板转子低速离心机3种方法制备细胞涂片,经瑞氏-姬姆萨染色后,显微镜下观察拍照。发现直接涂片中细胞图像难以观察,效果差,而细胞涂片离心机和酶标板转子低速离心机制备的细胞涂片,细胞图像清晰,均可达到细胞制片要求。酶标板转子低速离心细胞制片是一种便捷的新方法。     

花生胚乳细胞化的超微结构观察

花生（ＡｒａｃｈｉｓｈｙｐｏｇｅａｅＬ．）心形胚期的胚乳游离核多瓣裂,或具长尾状结构。胚乳细胞质内有大量线粒体、质体、高尔基体、小泡及少量内质网。中央细胞壁有壁内突。球胚及心形胚期常见胚乳瘤。心形胚晚期,胚乳开始细胞化,胚乳细胞壁形成有３种方式,分别存在于不同的胚珠中：（１）从胚囊壁产生自由生长壁形成初始垂周壁,具有明显的电子密度深的中层,其生长主要靠末端的高尔基体小泡及内质网囊泡的融合。两相邻的自由生长壁末端或其分枝末端相连形成胚乳细胞。（２）核有丝分裂后产生细胞板,细胞板向外扩展并可分枝。间期的非姊妹核间也观察到形成了细胞板。小泡与微管参与细胞板的扩展,高尔基体和内质网是小泡的主要来源。细胞板的扩展末端相互连接,形成胚乳细胞的前身。小泡继续加入细胞板的组成,以后形成胚乳细胞壁。（３）胚乳细胞质中,出现一些比较大的不规则形的片段性泡状结构,它们可能来源于高尔基体小泡,这些片段性泡状结构随机相连形成细胞壁,未见微管参与。胚乳细胞外切向壁及经向壁上有壁内突。     

Chloroplast Development in Rye Coleoptiles

In the parenchyma cells of 1-d-old dark-grown rye coleoptiles (Secale cereale) proplastids occurred which sometimes contained starch grains. During coleoptile growth in darkness starch-filled amyloplasts are formed from the preexisting proplastids. No prolamellar bodies were observed in the stroma of the plastids of the etiolated coleoptile. After irradiation of 3-d-old etiolated coleoptiles with continuous white light three different types of plastids occurred. In the epidermal cells proplastids were observed. The parenchyma cells below the stomata of the outer epidermis (above the two vascular bundles) contained mature, spindle-shaped chloroplasts with a well-developed thylakoid system. In the parenchyma cells that surround the vascular bundles amyloplasts with some thylakoid membranes (chloroamyloplasts) occurred. The mesophyll cells of the primary leaves of dark-grown seedlings contained etioplasts with large prolamellar bodies. In the primary leaves of irradiated plants chloroplasts similar to those of the parenchyma cells of the coleoptile were observed. Our results show that the rye coleoptile, which grows underground as a heterotrophic organ, is capable of developing mature chloroplasts upon reaching the light above the soil surface. The significance of this expression of photosynthetic capacity for the carbon economy of the developing seedling is discussed.     

The use of radioautography for investigating wall secretion in plant cells

Summary The paper summarises results of simple radioautographic experiments using tritiated glucoses to investigate wall secretion in plant cells. In outer root cap cells, labelled material was first concentrated in the Golgi bodies; it later appeared in vesicles, and was incorporated into the wall immediately under the plasmalemma. It finally collected mainly in the slime layer surrounding the root tip. Biochemical analyses have indicated that this material was pectic in nature. In inner root cap and epidermal cells, labelled material incorporated into the walls and also the cell plates of dividing cells was also apparently mainly derived from Golgi bodies. In meristematic (less differentiated) cells, however, the endoplasmic reticulum was more frequently labelled than the Golgi bodies near walls that had incorporated derivatives of labelled glucose. Considerable incorporation of labelled derivatives into the wall thickenings in coleoptile xylem cells was often detected; nearby elements of the endoplasmic reticulum were again frequently labelled in these cells and less often, Golgi bodies and the cytoplasm in the region occupied by microtubules contained radioactivity. Labelling of starch grains in the plastids was generally observed, but not in cells secreting large amounts of wall materials (outer root cap and older xylem cells); however, addition of larger amounts of exogenous glucose to outer root cap cells, following their incubation in tritiated glucose, promoted such incorporation. The paper finally sets forth some considerations on experimental techniques for radioautography that might be of more general application.     

Ultrastructure of palatal taste buds in the perihatching chick.

Palatal taste buds of perihatching chicks were examined by electron microscopy. Four intragemmal cell types were characterized. 1) Light: with voluminous, electron-lucent cytoplasm containing scattered free ribosomes, rough and smooth endoplasmic reticulum, plump mitochondria, sparse perinuclear filaments, occasional Golgi bodies, and numerous clear and dense-cored vesicles. Clear vesicles sometimes aggregate in a presynaptic-like configuration apposed to an axonal profile. These cells contained large, spherical, uniformly granular nuclei with one nucleolus. 2) Dark: with dense cytoplasm containing filamentous bundles surrounding the nucleus, occasional clear vesicles, centrioles, rough endoplasmic reticulum, and compact mitochrondria. The apical cytoplasm noticeably lacks dense secretory granules. Irregular to lobulated nuclei are densely granular, and contain scattered clumps of chromatin, adhering especially to the inner leaflet of the nuclear membrane, and at least one nucleolus. Cytoplasmic extensions of dark cells envelop other intragemmal cell types and nerve fibers. Light and dark cells project microvilli into the taste pore. 3) Intermediate: contain gradations of features of light and dark cells. 4) Basal: darker than the other intragemmal cell types and confined to the ventral bud region. Putative afferent synapses in relation to light cells, and axo-axonal contacts are described. While the appearance of axo-axonal contacts may be a transient developmental event, other bud features are consonant with observations in adult chickens and suggest that the peripheral gustatory apparatus is mature at hatching in this precocial avian species.     

Developmental changes in the inner epidermis of the bean seed coat

Summary The inner epidermis of the bean seed coat shows remarkable structural changes during seed development. At the globular stage of development, a moderately electron-dense substance begins to accumulate in the outer tangential and radial walls of the cells. The staining and fluorescence characteristics, together with the localization of peroxidase in the wall, suggest that this electron-dense material is a phenolic substance. At the same stage of embryo development, structural specialization can be detected in the cytoplasm of the epidermal cells with an increase in the abundance of organelles, especially the endoplasmic reticulum, mitochondria, and dictyosomes. These structural features are similar to those in the underlying branched parenchyma cells. As the seed rapidly expands during the maturation stage of embryo development, the epidermal cells and the inner layers of the branched parenchyma cells begin to degenerate. Small ruptures can be detected in the epidermis, exposing the branched parenchyma cells. These structural changes are discussed in relation to their possible functions during embryo development.     

ULTRASTRUCTURAL STUDIES OF PINUS PINASTER NEEDLES: THE ENDODERMIS

The endodermis in the needles of Pinus pinaster was examined with light and electron microscopy. The endodermis is composed of very long, radially flattened cells, filled with a large central vacuole, which contains spherical dense bodies whose concentration decreases from the ends of the cell to the middle part. They are individually surrounded by a fine granular matrix. The central vacuole is bounded by a thick tonoplast. Other small, clear vacuoles are limited by a thin tonoplast. The parietal cytoplasm contains relatively few ribosomes, long slender chloroplasts, and lipid bodies. The smooth endoplasmic reticulum is highly developed along the tangential walls and frequently connected, or apposed, to the plasma membrane. Numerous primary pit fields are seen in the radial walls which are lignified and in the tangential walls; the latter exhibit a characteristic loosening of the outer layer of the wall. The lipid bodies are connected to endoplasmic reticulum tubules. The role of the endodermis in the active transport of water inside the needle is discussed in reference to previous physiological studies. The chemical composition of the vacuolar dense bodies is as yet unknown.     

The ultrastructural development of the macrogamete ofEimeria stiedai

Summary The development of the macrogamete ofEimeria. stiedai in the epithelial cells of the bile ducts of rabbits was studied by electron microscopy. A macrogamete was first identified by the presence of a large central nucleus with prominent nucleolus, and subsequently by the appearance of wall forming bodies. The macrogamete was limited by an outer single membrane under which there were remnants of a second membrane. The parasitophorous vacuole, in which the macrogamete was located, was often narrow and it contained no intravacuolar-tubules or -folds. As macrogametogony proceeded wall forming bodies of Type I and II, canaliculi, electron pale spaces (lipid) and polysaccharide granules increased in number. Granular endoplasmic reticulum, mitochondria and Golgi bodies were present throughout.     

Ultrastructure of lineus ruber (rhyncocoela) epidermis

Recent evidence indicates that nemertean epidermis is capable of absorbing certain organic solutes from sea water via mediated transport mechanisms, as well as secreting mucoid substances. Morphological studies suggest that these functions may be restricted to distinct epidermal cell populations.Mucous secretion at the free surface of the epidermis is the result of synthesis and release activities of cells in both the epidermis and dermis (cutis). Secretion of dermal origin passes through the epidermis to the worm's exterior in slender cytoplasmic processes (canaux d'evacuation) in the form of membrane bound vesicles. A single gland cell type, located entirely within the epidermis, releases externally a granular product histochemically identified as largely protein plus some amount of carbohydrate with low periodic acid-Schiff's reactivity. The close juxtaposition of granular endoplasmic reticulum and Golgi apparati to the secretory material is consistent with the composition of this secretory product.Interstitial cells possess microvilli projecting from their apical surface, in addition to cilia. The outer surface of the plasmalemma covering these ciliary projections is unadorned, but microvilli possess a fuzzy coat. At the peripheral ends of the microvilli, the coat is filamentous, while at their base the coat consists of foliate structures. Cationic colloidal iron binding suggest that the filamentous portion of the fuzzy coat contains the greatest proportion of the acidic surface charge. The presence of periodic acid-Schiff's positive material in this region suggests that the fuzzy coat also contains carbohydrate. Lateral boundaries of the interstitial cell lacks obvious junctional specializations; however, the apical 150 nm intracellular space narrows to 40 nm and continues in a tortuous interdigitating path to the base of the adjacent interstitial cells. Where the apex of these cells is broad, the interdigitations are shallow, but the basal half of the interstitial cells have deep complex infoldings. Cytoplasmic organelles other than the nucleus, mitochondria and some granular endoplasmic reticulum, are restricted to the apical half of the cytoplasm. The presence of closely apposed Golgi complexes and smooth endoplasmic reticulum, multivesicular bodies, lysosome-like dense vesicles and coated vesicles suggests that these cells may play a role in intracellular digestion of phagocytized paniculate matter from the external environment. The amplification of the interstitial cell's free surface suggests that these cells are primarily responsible for mediated solute transport across the epidermis.     

A new set of vesicles in Giardia lamblia

In the present study it was demonstrated the existence of a new set of membrane-bounded vesicles in Giardia lamblia. They were found in dividing and non-dividing trophozoites studied by routine transmission electron microscopy, freeze-fracture and Thiéry's technique. Encysting cells were not studied. These vesicles appear different to the previously reported components of the Giardia endomembranous system, such as the endoplasmic reticulum (ER), lysosome-like peripheral vesicles (PV), and the encystation-specific vesicles (ESV) that appear during trophozoite differentiation into cysts. They measure 100-150 nm in diameter, and thus are smaller than the peripheral vesicles, and the encystation-specific vesicles (ESV). They were found in clusters, scattered throughout the cytoplasm, but preferentially located close to the nuclei, axonemes, median bodies, and ER profiles. These internal vesicles are roughly spherical, and their contents present different electron densities and are more electrondense than those of the peripheral vesicles. They appeared to be budding from the outer nuclear membrane envelope. These cytoplasmic vesicles were found only in cells with very good fixation. Only few cells in the same preparation exhibited these vesicles.     

Localization of lipocaline-type prostaglandin D synthase in rat brain: immunoelectron microscopic study

The distribution of lipocaline-type prostaglandin D synthase (L-PGDS) in rat brain was investigated by immunoelectron microscopy using a protein A-gold technique. In perivascular cells adjacent to the basement membrane of arterioles in the pia-arachnoid and of blood vessels in the subpial cortex, gold labeling was confined to the lumen of the dilated rough endoplasmic reticulum, and not found in the few lysosomes present in the cytoplasm. The results suggest that the perivascular cells secrete L-PGDS and seem not to degrade lipophilic molecules carried by L-PGDS. Moreover, gold particles representing the antigenic sites of L-PGDS were found in the Golgi apparatus, rough endoplasmic reticulum, vesicles, and nuclear envelope of arachnoid trabecular cells, arachnoid barrier cells, and arachnoid pia mater cells. The labeling was less detectable in the same organelles of choroid plexus epithelial cells, compared with leptomeningeal cells. In meningeal macrophages and parenchymal microglia, L-PGDS was detected in lysosomes, multivesicular bodies, and endocytic vesicles. The production of L-PGDS in perivascular cells is important to the various functions of this enzyme in brain parenchyma.