一种制备植物扫描电镜样品的新干燥法：叔丁醇冻结干燥法

叔丁醇冻结干燥法用于植物材料,获得了良好的效果。经常规固定的样品完成脱水以后浸泡在叔丁醇中,然后将含醇样品放入电冰箱中冷冻,待叔丁醇凝固后再转移到真空镀膜机的钟罩里,用机械泵抽真空,冻结的醇在低真空中升华后样品得到了干燥。SEM 镜检表明,样品的表面和细胞断裂面及其深层(可见到内质网的双层膜、线粒体的嵴、高尔基体潴泡叠层、核糖体颗粒等细胞器)的三维图象均优于用临界点干燥法所得到的样品图象,叔丁醇冻结干燥法用于植物 SEM 样品制备,是一种操作简单,干燥质量可靠、值得推广的方法。     

植物细胞壁富含羟脯氨酸糖蛋白（HRGP）的电子显微镜观察

本文介绍了一种简单可行的,用以观察和鉴定一种植物细胞壁蛋白质－富含羟脯氨酸糖蛋白（HRGP）的方法,胡萝卜HRGP属于一种伸展蛋白（extensin)。通过简化的装置将HRGP喷射到云母片上,再经旋转投影,使其在透射电子显微镜下呈现为棒状分子（rod-like molecule),经电镜观察和测量表明,HRGP的分子长度为87nm。     

葫芦科8属11种植物花粉形态的扫描电镜观察

利用扫描电子显微镜,对葫芦科8属10种1变种植物的花粉形态进行观察研究。结果显示:冬瓜属、葫芦属和栝楼属的花粉粒为近球形,具3孔沟;苦瓜属、西瓜属和丝瓜属的花粉粒为长球形,具3沟;黄瓜属的花粉粒为近球形,具3孔;南瓜属的花粉粒为球形,具散孔。花粉粒大小、形状和外壁雕纹属、种间差异显著。     

高分子量激肽原富含组氨酸区域抑制细胞伸展的机制分析

活化型高分子量激肽原 (activehighmolecularweightkininogen ,HKa)是组织培养板上体外连接蛋白 (vitronectin ,VN)促使细胞伸展的潜在抑制物 ,已证实轻链的富含组氨酸区域 (histidine richdomain ,HRD)是HKa抗细胞伸展的活性区域 .HK的重组HRD (r HRD)能够促使成纤维细胞伸展 .通过基于HRD序列的选择肽分析 ,定位了HRD的细胞伸展序列 .5个肽中的 3个能够使TIG 3细胞伸展 .P 1肽引起的细胞伸展能够被可溶性P 5肽或HKa所抑制 .P 2肽不能抑制P 1或P 5肽引起的细胞伸展 .r HRD以及 3种肽介导的细胞伸展能够被RGD合成肽以及抗αvβ3或α5β1整合素抗体所抑制 .结果提示 ,选择肽引起的细胞伸展是由整合素介导的 ,尽管此区域不含有RGD序列     

三种石杉属植物孢子形态的扫描电镜观察

利用扫描电镜对采自湖北省利川市的蛇足石杉Huperzia serrata（Thunb.ex Murray）Trev.、皱边石杉H.crispate（Ching ex H.S.Kung）Ching和四川石杉H.sutchueniana（Herter）Ching3种植物孢子的大小、形态及表面纹饰等方面进行观察测量,并对每个种孢子形态特征进行了描述。结果表明,3种石杉属植物孢子均为四面体型、三裂缝、辐射对称,表面纹饰均呈穴状。不同种在孢子大小、裂缝长度、孔穴深浅程度以及辐射间区凹陷程度上存在差异。因孢子形态是稳定的,可作为种间划分的重要依据,同时,也为蕨类植物孢子形态学的研究提供参考。     

蜘蛛抱蛋属植物叶表皮微形态的扫描电镜观察

在扫描电子显微镜下,观察9种蜘蛛抱蛋属植物的叶表皮微形态。结果表明,9种植物叶下表皮的细胞都为长方形,垂直壁为直的,垂周界限为凸的,外层周壁凹陷。气孔椭圆形至卵圆形,气孔外拱盖表面多平滑,其内缘近平滑、浅波状或锯齿波状;角质膜多为脊状条纹、有的外层周壁有横纹或颗粒。气孔大小与染色体的倍性有一定的正相关关系。不同种间叶表皮特征表现出一定差异,对种的划分有一定的分类鉴定意义。     

高淀粉膳食对血浆胰岛素、cAMP含量及组织cAMP代谢的影响

对高淀粉膳食(糖占总热量80％)对大鼠血脂、胰岛素及cAMP代谢的影响进行了研究。大鼠摄取高淀粉膳食3天,空腹血浆岛素及甘油三酯含量明显高于对照组(P<0.01;P<0.01)。6天后血浆甘油三酯含量增高近四倍(P<0.01),而血浆、肌肉和脂肪组织cAMP含量低于对照组,分别减低38％,45％和32％(P<0.05;P<0.05,0.1相似文献   

玉米灌浆期水分差异供应对籽粒淀粉积累及其酶活性的影响

利用普通玉米(Zay mays)‘掖单22’和高油玉米‘高油115’,研究了灌浆期水分差异供应对籽粒淀粉及其组分积累、相关酶活性动态变化的影响。结果表明,两种类型玉米淀粉积累和酶活性动态变化趋势基本一致,但对水分的反应有差异。缺水提高了‘掖单22’籽粒中淀粉、支链淀粉含量,而直链淀粉含量下降,‘高油115’则是籽粒中的淀粉含量、支链淀粉和直链淀粉含量提高;充分供水使淀粉及其组分产量提高;叶片中蔗糖合成酶(SS)、磷酸蔗糖合成酶(SPS)活性随水分供应水平而提高,尤其在授粉后10~30 d增幅更加明显。充分供水明显提高籽粒中腺苷二磷酸葡萄糖焦磷酸化酶(ADPG-PPase)、尿苷二磷酸葡萄糖焦磷酸化酶(UDPG-PPase)、可溶性淀粉合成酶(SSS)和淀粉粒结合淀粉合成酶(GBSS)活性,缺水使籽粒中酶活性下降较早且迅速;SPS、ADPG-PPase、SSS酶活性对缺水反应比较敏感。     

水分胁迫下小麦叶片蛋白质、淀粉和纤维素合成的示踪研究

利用光合１４ＣＯ２示踪技术,在－０．８ＭＰａＰＥＧ处理下,对等基因型不同抗旱性ＫＴＣ８６２１、ＮＤ７５３２和本地抗旱品种陕合６号小麦进行比较,其叶组织细胞质膜相对透性增加２７％－３２％,合成蛋白质放射性下降６２％－７０％;合成淀粉放射性下降必３％－６９％;合成纤维素放射性下降５９％－７１％。其中高抗旱的ＫＴＣ８６２１１大分子合成下降幅度小于等基因型低抗旱的ＮＤ７５３２和陕合６号,表明该品种叶体内大分子合成受水分胁迫的影响较小。正常水分条件下等基因型两品种叶酶溶物放射性及在ＰＥＧ干旱处理下的下降幅度均相近。陕合６号叶醇溶物放射性远远小于前两品种．     

高油、高淀粉玉米吸硫特性及施硫对其产量、品质的影响

研究了高油、高淀粉玉米硫素的吸收分配及施硫对籽粒产量、品质的作用。结果表明：在11250kg/hm^2的产量水平下,开花前硫素吸收量占总吸收量的百分比平均为57.53%,花后对硫的吸收较强;高油1号、长单26、掖单13每公顷吸硫量分别为：32.46kg、34.94kg,34.20kg。施硫能增加穗粒数和粒重（高油玉米粒重增加不显著）,并显著提高其产量,施硫22.5kg/hm^2（S1）能使籽粒含油率和蛋白质含量分别比对照提高9.05%和6.88%,蛋白质中清蛋白、球蛋白和谷蛋白含量提高,品质改善;施硫量增至90kg/hm^2(S2)时,籽粒含油率和蛋白质含量仍有提高,但蛋白质中醇溶蛋白含量增加,蛋白质品质降低。     

Drying techniques for the visualisation of agarose‐based chromatography media by scanning electron microscopy

The drying of chromatography resins prior to scanning electron microscopy is critical to image resolution and hence understanding of the bead structure at sub‐micron level. Achieving suitable drying conditions is especially important with agarose‐based chromatography resins, as over‐drying may cause artefact formation, bead damage and alterations to ultrastructural properties; and under‐drying does not provide sufficient resolution for visualization under SEM. This paper compares and contrasts the effects of two drying techniques, critical point drying and freeze drying, on the morphology of two agarose based resins (MabSelect?/d _w ≈85 µm and Capto? Adhere/d _w ≈75 µm) and provides a complete method for both. The results show that critical point drying provides better drying and subsequently clearer ultrastructural visualization of both resins under SEM. Under this protocol both the polymer fibers (thickness ≈20 nm) and the pore sizes (diameter ≈100 nm) are clearly visible. Freeze drying is shown to cause bead damage to both resins, but to different extents. MabSelect resin encounters extensive bead fragmentation, whilst Capto Adhere resin undergoes partial bead disintegration, corresponding with the greater extent of agarose crosslinking and strength of this resin. While freeze drying appears to be the less favorable option for ultrastructural visualization of chromatography resin, it should be noted that the extent of fracturing caused by the freeze drying process may provide some insight into the mechanical properties of agarose‐based chromatography media.     

A benign approach to the preparation of freshwater bryozoan statoblasts for scanning electron microscopy (SEM) imaging

Abstract

Several different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanella and Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals.     

A Chemical Drying Technique for Plant Materials Applied to the Scanning Electron Microscopy

There are several drying methods for biological materials for the use of scanning electron microscopy. Applying bexamethyl disilazane (HMDS) as a drying treatment is a new method and it's application on drying plant tissue has not been previously reported. The advantage of this method is the treatment only for a few minutes and is also good and stable for very small biological specimens. The method is simple, low cost and time saving and does not need any apparatus. The features of the tissue structure observed are satisfactory.     

扫描电子显微镜植物材料的一种化学干燥技术

扫描电镜新鲜生物材料的制备,一般都要经过干燥处理,其目的主要是使样品不产生收缩畸变,以使样品的形态结构保持在生活状态,如此获得的样品图像才自然真实。现代广为应用的扫描电镜生物样品的干燥方法是临界点干燥(CPD)法。另外还有莰烯干燥法、乙腈干燥法和叔丁醇干燥法等。Jemes(1983)应用化学试剂六甲基二硅胺烷(nexamthyl disilazane,简称 HMDS),分子式[(CH_3)_3Si]_2对昆虫组织进行干燥处理并     

Scanning electron microscopy preparation protocol for differentiated stem cells

The lack of an established protocol for scanning electron microscopy (SEM) studies on stem cells differentiating into adipogenic lineage led us to develop a protocol for the preparation of differentiated adult bone marrow-derived mesenchymal stem cells (BMSC) for SEM. This protocol describes the procedure to maintain and preserve the structural organization of cellular components following differentiation, for morphological and physical characterization. The fixation of the differentiated cells was followed by dehydration using methanol, and vacuum desiccation before microscopy. The use of longer chain alcohols as dehydrating agents was avoided in our method to reduce the dissolution of lipid deposits in cells, thus allowing the maintenance of their structural integrity. The time period for the processing of samples was reduced by avoiding the osmium tetroxide postfixation and critical point drying. Thus, this protocol helps in determining the potential, fate, and degree of stem cell differentiation. This may be useful for SEM analysis of differentiated cells, especially those grown on various scaffolds.     

A replication technique for scanning electron microscopy: Applications for anthropologists

Scanning electron microscopy has become an increasingly useful tool for anthropologists, particularly because of the development of improved methods of replicating specimens. One of the best replication techniques involves silicone-based dental impression materials to make negative impressions, in conjunction with epoxy resins, which are used to make positives or casts. The technique outlined here is particularly useful for anthropologists. Using this technique allows the examination of bone surfaces, teeth, and fossils for taphonomic, microwear, and experimental studies. Reproduction of detail is faithful at magnifications of × 1,500 to × 2,000, routinely giving resolutions of .1 to .25 μm.     

培养破骨细胞扫描电镜样本制备方法的探讨

目的：探讨体外培养的破骨细胞在自制牛股骨磨片和细胞爬片中扫描电镜制备方法。方法：实验分两组,一组采用新鲜牛股骨制备成5mm×5mm大小的薄片,作为共培养之需;另一组,采用盖玻片制成5mm×5mm的细胞爬片。分别以5×104种植于骨磨片和爬片,培养5天后进行扫描电镜的制备并观察。结果：破骨细胞在牛骨磨片表面生长良好,充分伸展,有细胞突起伸入到实验组材料深部,并形成骨陷窝;在爬片表面生长的破骨细胞,细胞生长良好,粘附性强,细胞之间相互连接较紧密,细胞表面突起明显。结论：牛股骨磨片与破骨细胞在体外相容良好,材料有利于破骨细胞的生长及细胞功能的表达,而破骨细胞爬片更适于细胞外形的观察。将两种方法结合既能反映破骨细胞的形态结构又能展示其破骨功能。     

培养破骨细胞扫描电镜样本制备方法的探讨

目的:探讨体外培养的破骨细胞在自制牛股骨磨片和细胞爬片中扫描电镜制备方法。方法:实验分两组,一组采用新鲜牛股骨制备成5mm×5mm大小的薄片,作为共培养之需;另一组,采用盖玻片制成5mm×5mm的细胞爬片。分别以5×104种植于骨磨片和爬片,培养5天后进行扫描电镜的制备并观察。结果:破骨细胞在牛骨磨片表面生长良好,充分伸展,有细胞突起伸入到实验组材料深部,并形成骨陷窝;在爬片表面生长的破骨细胞,细胞生长良好,粘附性强,细胞之间相互连接较紧密,细胞表面突起明显。结论:牛股骨磨片与破骨细胞在体外相容良好,材料有利于破骨细胞的生长及细胞功能的表达,而破骨细胞爬片更适于细胞外形的观察。将两种方法结合既能反映破骨细胞的形态结构又能展示其破骨功能。     

Giardia muris: scanning electron microscopy of in vitro excystation

A recently developed in vitro excystation procedure results in almost total excystation of Giardia muris, an intestinal parasite of mice. The present experiment examines the G. muris cyst morphology by scanning electron microscopy and the efficacy of the excystation procedure. Untreated cysts of G. muris were elliptical and displayed a distinctive surface structure. Excystation began almost immediately after incubation had begun and most trophozoites emerged within 30 min. Excystation appears to involve flagellar action of the encysted trophozoite. A tear of the wall occurred at one pole. This opening was subsequently enlarged, presumably by flagellar action. Trophozoites emerged, posterior end first, and an associated mucoid-like material was extruded. Newly emerged trophozoites were nearly oval in shape. Trophozoites quickly became flattened, elongate, and underwent cytokinesis resulting in two daughter trophozoites. Few organisms not excysted were seen after 30 min incubation.     

Aspects of the three-dimensional intracellular organization of mesocarp cells as revealed by scanning electron microscopy

Summary The osmium-ligand binding technique and scanning electron microscopy have been applied to the study of the three-dimensional organization of mesocarp cells of a mature avocado fruit. Using this approach the mitochondria of the cells appear as elongated, branching structures and the endoplasmic reticulum consists of a complex of tubular strands, vesiculated strands and lamellar sheets. Associations of the endoplasmic reticulum with other organelles are also apparent. It is suggested that this approach provides a valuable means to assess the structural transitions in cell organization that occur during development or with functional changes.