The quest for four-dimensional imaging in plant cell biology: it's just a matter of time

BACKGROUND: Analysis of plant cell dynamics over time, or four-dimensional imaging (4-DI), represents a major goal of plant science. The ability to resolve structures in the third dimension within the cell or tissue during developmental events or in response to environmental or experimental stresses (i.e. 4-DI) is critical to our understanding of gene expression, post-expression modulations of macromolecules and sub-cellular system interactions. SCOPE: Microscopy-based technologies have been profoundly integral to this type of investigation, and new and refined microscopy technologies now allow for the visualization of cell dynamics with unprecedented resolution, contrast and experimental versatility. However, certain realities of light and electron microscopy, choice of specimen and specimen preparation techniques limit the scope of readily attaining 4-DI. Today, the plant microscopist must use a combinatorial strategy whereby multiple microscopy-based investigations are used. Modern fluorescence, confocal laser scanning, transmission electron and scanning electron microscopy provide effective conduits for synthesizing data detailing live cell dynamics and highly resolved snapshots of specific cell structures that will ultimately lead to 4-DI. This review provides a synopsis of such technologies available.     

Cellular architecture and migration behavior of fibroblast cells on polyhydroxyoctanoate (PHO): A natural polymer of bacterial origin

Biodegradable and biocompatible novel materials of natural origin are gaining more and more attention in recent years. These so called biopolymers, characterized by their biointegrity and biocompatibility, find completely new and promising applications in biomedical sciences. The presented work focuses on the medium chain length elastomeric polyhydroxyalkanoate biopolymer—polyhydroxyoctanoate (PHO). This biopolymer is fully biodegradable without formation of harmful byproducts.We investigated PHO's physical properties with nanoindentation technique and scratch testing to determine Young's modulus and friction coefficient. Further, the work focused on the impact of PHO, used as growth substrate, on the physiology and morphology of mouse embryonic fibroblast cells (MEF 3T3). Application of fluorescent staining protocols and advanced microscopic techniques allowed to study the morphological changes in the cytoskeletons of cells grown on PHO and also gave an insight into their migration strategies on the polymer surface. We found that PHO exhibits no cellular cytotoxicity, similarly to a glass substrate. MEF cells spread better on glass surface than on each tested PHO substrate though there was almost no difference between PHO substrates cast from different solvents. However, a detailed analysis of actin and microtubule cytoskeletal architecture reveals changes in the density of actin and microtubular networks. Migration of MEF cells on PHO substrates was slower than on the glass substrate. To elucidate the molecular mechanisms of observed changes in cytoskeletal architecture and migration parameters can be of special interest for future medical application of PHO polymer.     

黑龙江椴树属植物叶表皮形态学观察

利用扫描电子显微镜对紫椴（Tilia amurensis Rupr.）及糠椴（T. mandshurica Rupr.et Maxim.）叶片的微观形态结构特征进行观察,并进行微观形态结构的比较研究。研究结果表明紫椴与糠椴叶片上表皮微观形态相同,而下表皮气孔器的形态类型、蜡质纹饰和角质膜的形态特征、单位面积上星状毛的数量均表现出很大的差异。这些微观形态差异可作为区分紫椴和糠椴的分类学依据。     

动态检测血管紧张素Ⅱ刺激后STAT3信号核转位及其与衰老的关系

为了动态观察复制性衰老细胞中血管紧张素Ⅱ(AngⅡ)激活人信号转导与转录活化因子3(hSTAT3)信号转导途径的核转位情况及该途径在细胞衰老过程中的变化。将载体pMS1-hSTAT3上的目的基因STAT3序列亚克隆到pEGFP-C3报告载体中,构建出pEGFP-hSTAT3质粒选择人胚肺二倍体成纤维细胞WI-38细胞株进行细胞培养,通过脂质体(Effectene)转染的方法,将pEGFP-STAT3质粒分别转染至19代,42代WI-38细胞中;在激光共聚焦显微镜下动态观察血管紧张素Ⅱ激活STAT3的核转位的变化及不同代数的差别．结果显示AngⅡ刺激细胞后,出现活化的STAT3在胞核内聚集现象,并且在19代细胞中15分钟开始出现,30—45分钟左右时达到高峰;42代细胞中30分钟左右开始出现,50—60分钟左右达到高峰。因此我们认为AngⅡ刺激WI-38细胞,能出现hSTAT3信号转导的核内集聚现象;并且随着WI-38细胞传代数的增加,STAT3信号转导入核时间延迟。说明随着细胞的复制性衰老,通过STAT3通路转导的信号活性逐渐降低,最终影响细胞的增殖,可能是促进细胞衰老的原因之一。     

Antinematodal activity and the mechanism of the antimicrobial peptide, HP (2-20), against Caenorhabditis elegans

The peptide HP (2-20), derived from the N-terminal sequence of Helicobacter pylori ribosomal protein L1 (RPL1), has a nematicidal activity against eggs and worms of Caenorhabditis elegans. Eggs treated with HP (2-20) (69%) has a higher fluorescence intensity with propidium iodide staining, which was similar to that of melittin (82%) but higher than untreated cells (5.7%). Confocal microscopy showed that the peptides were located in the shell of the eggs and the inner and outer surfaces of the worms. HP (2-20) therefore may exert its antinematodal activity by disrupting the structure of the egg's shell and the cell membrane via pore formation or by direct interaction with the lipid bilayers in a detergent-like manner.     

Identification of hairs found in leopard seal (<Emphasis Type="Italic">Hydrurga leptonyx</Emphasis>) scats

The leopard seal is a top-order predator in the Southern Ocean ecosystem and preys on a wide variety of vertebrate species including seals and penguins. We assessed the use of hairs found in leopard seal scats to identify the species of pinniped consumed. A reference collection of hairs was obtained from four potential leopard seal prey species including crabeater, Weddell, Ross, and Southern elephant seals. Discrimination techniques applied to terrestrial mammals did not allow for identification of the seal hairs. Instead, a 2-dimensional (2-D) and 6-dimensional (6-D) analysis technique utilising Mahalanobis distances (D ²) was used. The smallest Mahalanobis distance together with the largest value of p(F) positively identified hairs from each species. The 6-D analysis was more accurate and applied to hairs found in the leopard seal scats. The majority of prey species were identified as crabeater seals, which are a known prey item of the leopard seal.     

An examination of Stone Age/Bronze Age adzes and axes of red deer (<Emphasis Type="Italic">Cervus elaphus</Emphasis> L.) antler from the Leine Valley,near Hannover

Red deer antler was, together with stone, one of the most important raw materials for tool manufacture in prehistoric times. Both the physical characteristics of this material and the manufacturing technology of antler tools are examined. The use of experimental tests and analytical studies of the recovered antler finds allow us to propose, with a great degree of confidence, that the main use of antler axes during the Stone Age/Bronze Age was in woodworking.     

多聚亚基蛋白质在固-液界面团聚的测量与表征

以血红蛋白和乙肝表面抗原(HBsAg)为模型,联用双偏振光干涉分析仪(DPI)和原子力显微镜(AFM)研究在固-液界面上的吸附行为.结果发现:当溶液环境中血红蛋白的质量浓度为1.08 mg/mL时,测得在硅羟基表面吸附的平均厚度为3.54 nm,蛋白颗粒直径达60～ 150 nm,远高于血红蛋白的分子尺寸,说明血红蛋白在固-液界面上发生团聚.当HBsAg质量浓度为0.67 mg/mL时,测得在硅羟基表面及DEAE琼脂糖表面吸附的平均厚度分别为1.06和23.69 nm,表面吸附的蛋白颗粒直径分别为288～401和520 ～ 971 nm,是单分子HBsAg尺寸的13.1～44.1倍,表明在模型蛋白表面都形成了大的团聚体.     

大鳞副泥鳅鳞片表面结构的扫描电镜观察

对大鳞副泥鳅的鳞片作了扫描电镜观察。结果表明,大鳞副泥鳅具圆鳞;基区、侧区和顶区均具有辐射沟及环沟,二者交织成网状,将鳞片分割成块状,可增加鳞片的柔软度;次级辐射沟发出部位可作为确定年轮的主要依据。     

紫彩血蛤鳃的组织化学和扫描电镜研究

紫彩血蛤（Ｎｕｔｔａｌｌｉａｏｌｉｖａｃｅａ）的鳃上皮细胞核多为椭圆形,核内染色体呈丝状;结缔组织细胞的核多为圆形,部分核内有块状Ｆｅｕｌｇｅｎ阳性颗粒,ＲＮＡ在鳃丝上皮细胞最丰富,有的形成直径０．５２μｍ的颗粒,结缔组织和鳃丝上皮细胞内均有直径０．６３μｍ左右的ＰＡＳ阳性颗粒,吞噬细胞呈极强的ＡＮＡＥ阳性,鳃丝上皮细胞呈较弱的阳性。ＡＣＰ和ＡＫＰ在吞噬细胞和外缘上皮细胞中的活性较强,其它部位较弱