黄芪的胚胎学研究Ⅰ、雌雄配子体发育

通过光学显微镜对黄芪雌雄配子体的发育过程进行观察, 同时对花粉发育进行细胞化学实验, 其主要结果如下;1.花粉第一次有丝分裂形成一个较小的半球形生殖细胞和一个较大的营养细胞。2.生殖细胞发育过程中存在暂短的细胞壁, 经PAS反应和苯胺兰荧光显微反应鉴定均为负反应。即:生殖细胞壁并未显示出纤维素或胼胝质性质的壁。3.营养细胞与生殖细胞之间壁的解体及生殖细胞进入营养细胞的过程。4.成熟花粉中的生殖核为孚尔根反应强阳性, 营养核为弱的正反应。5.雌配子体发育起源于珠心组织亚表皮下的孢原细胞其中只有一个孢原直接发育成为蓼型胚囊。 文中对黄芪花蕾外部形态及其内部雌雄配子体的发育作了相关性比较。     

川百合花粉管的生殖细胞分裂过程中微管骨架的分布变化

应用透射电镜辅以免疫荧光定位技术研究了川百合 (Lilium davidii Duch.)花粉管中生殖细胞分裂过程中染色体动态和微管分布的关系。在生殖细胞分裂前和有丝分裂前期 ,电镜观察一直未见微管结构 ,但免疫荧光图象显示生殖细胞中有微管蛋白存在。直到分裂的前中期—中期 ,染色体出现 ,它们沿花粉管的长轴前后排列 ,横向的着丝点对相应地一对对地纵向排列。这时 ,生殖细胞中才出现大量微管 ,它们分布于细胞周质区和染色体之间 ,并跨越染色体的整个长度。前中期—中期开始时 ,只有 1～ 2对着丝点从横向转为纵向 ,微管垂直插入着丝点形成着丝点微管 ,而非前人用免疫荧光方法观察到的微管与着丝点侧向联接的图象。随着横向的着丝点对逐渐转变成纵向的过程 ,着丝点微管数量逐渐增多 ,但不形成典型的纺锤体。分裂后期 ,染色体交错分离 ,微管的分布与前中期—中期的基本相同。晚后期 ,染色体呈明显的两群 ,除极区和细胞中央区有微管残余外 ,大部分微管消失。通过染色体长度的测量 ,间接证明了分裂后期 B的存在。分裂末期的晚期 ,核膜形成后 ,在两精核之间的区域 ,微管数量开始增多。此区可能代表用免疫荧光所观察到的微管重叠区。细胞板出现后 ,微管消失     

川百合精细胞的超微结构(英文)

川百合与朱顶红花粉管中的生殖细胞分裂行为非常不同。诸如：染色体行为、微管的组织形式和分布、包括着丝点、微管形成的时间,纺锤体的形状及间期周质微管网络在生殖细胞分裂过程中消失与否等。但这两种细胞具有某些共性,包括在有丝分裂前期缺乏早前期带微管（ＰＰＢ）,末期形成细胞板等。这两种植物精细胞的结构应有较大差异。我们曾报道了朱顶红精细胞的超微结构,本文详细从超微结构方面描述了川百合精细胞的特征。川百合花粉管的萌发采用半离体活体培养方式。１１～１８小时后,ＤＮＡ荧光染料Ｈｏｅｃｈｓｔ３３２５８和醋酸地衣红染色检查花粉管中生殖细胞和精细胞发育时期。切取含有分裂的生殖细胞和精细胞的花柱部分,按曾报道的方法固定、包埋、切片、染色及观察。在所有检查的花粉管中,两精子均前后排列（Ｆｉｇ．１～３）,营养核前导并靠近花粉管顶端（Ｆｉｇ．２,３）。Ｈ３３２５８染色可见两精核间以ＤＮＡ联系（Ｆｉｇ．３）。两个新形成的精核彼此分离（Ｆｉｇ．１）,后来又相互靠近,并维持一定距离（Ｆｉｇ．３）。偶尔一对精子与营养核靠近（Ｆｉｇ．２）。两精细胞被一共同的细胞壁连接,他们不仅被自己的质膜也被营养细胞的质膜包围构成周质。周质平坦光滑。共同壁横向     

问题解答

问植物细胞中的高尔基体跟细胞壁的形成有什么关系?(高中《生物》第22页)答植物细胞壁的形成,开始于细胞分裂后期末或末期初。在两组分离的染色体之间,分裂母细胞的赤道面上出现了新的微管,并形成了成膜体。这些成膜体微管在细胞板形成时供运输物质和移动小泡用。与此同时,在这个区域出现了许多来自高尔基体的小泡,小     

大蒜根尖细胞微管的免疫荧光观察

1963年,先后在动物和高等植物细胞中发现微管结构。已经知道微管不仅具有支持功能,而且在运动、运输和分泌等一系列细胞活动中发挥重要作用。在高等植物细胞中,微管明显地参与形态建成。周质微管(Cortical microtubules)可能与细胞壁中纤维素微纤丝的排列与定向有关。早前期带(Preprophase Bands)预示胞质分裂时细胞板的位置。成膜体微管参     

水平回转对水稻幼苗叶细胞的影响

对在模拟微重力装置上回转１４ 天的水稻幼苗叶细胞进行了亚显微形态、电子探针和细胞酶化学研究。发现叶细胞质膜上Ｃａ２＋ －ＡＴＰ酶活性消失,膜内钙总量上升、膜外钙总量下降,细胞骨架变得疏松,细胞壁变薄并凹凸不平。叶绿体的基粒和线粒体的内嵴亦有部分变化。其变化机制,首先是细胞质膜上Ｃａ２＋ －ＡＴＰ酶活性消失,膜上钙泵停止工作,跨膜钙浓度差减小,膜内钙浓度上升,微管、微丝聚合受阻,细胞骨架疏松,分泌泡移动失去导向,从而导致细胞壁变薄等状态     

小麦雄配子体的超微结构 Ⅰ.营养细胞和生殖细胞的形成

小孢子分裂的末期产生的成膜体,经离心的扩展后形成一个与内壁连结的细胞板,而后形成分隔营养细胞和生殖细胞的壁。由于胞质分裂高度的不均等性,形成大小悬殊的营养细胞和生殖细胞。在初期的生殖细胞和营养细胞的细胞质中,细胞器是没有差异的,包括线粒体、质体、内质网、高尔基体和核糖体。只有在胞质分裂初期看到微管。生殖细胞形成后,进一步的发育是逐渐脱离花粉粒的壁而成为游离的细胞,浸没在营养细胞的细胞质中。与此同时壁物质消失,变为一个被二层质膜所包围的裸细胞。当生殖细胞发育至游离的裸细胞时期,与营养细胞比较,显示明显的异质性,表现为生殖细胞中的质体不发育或退化,其它细胞器没有什么变化。相反,营养细胞中的质体和线粒体在数量上和大小上显著增长,在质体中迅速积累淀粉。对小麦生殖细胞暂时出现细胞壁的意义以及和营养细胞的异质性进行了讨论。     

小麦雄配子体的超微结构 Ⅱ.精细胞的形成和发育

1.通过小孢子有丝分裂而形成的生殖细胞,在发育过程中有一系列的包括位移和变形的变化。最后生殖细胞变为纺锤体,准备另一次有丝分裂。在此时期,生殖细胞是裸露的,它只有自己的质膜和被营养细胞的质膜包围。生殖细胞的大部分为明显的椭圆形的核所占据,具高度凝集的染色质。在生殖细胞薄层的细胞质中,除核糖体外,所有的细胞器比营养细胞质中的明显的少,而且小。微管也能在纺锤形的生殖细胞的细胞质中看到,它们的排列方向是和细胞的长轴平行的。在生殖细胞中没有造粉质体。2.当生殖细胞已移位并和营养细胞再次紧密靠近后,它开始分裂。由生殖细胞分裂所形成的精细胞及其发育包括下列主要的变化:精细胞的形状由圆球形变为椭圆形,最后变为具尾延长的细胞。与此同时,细胞质的分布逐渐集中到精细胞的一端,形成细胞质的延伸,构成所谓的精细胞的尾部。有更多的细胞器,特别是线粒体集中在尾部。从生殖细胞分裂刚形成的精细胞是裸露的,所包围的质膜是不连续的。除精细胞的质膜之外,为营养细胞的质膜所包围,在后来的发育时期此双质膜变为连续的,并在一个极短的时期有壁物质——胼胝质沉积在质膜之间的空间。但在此时期之后细胞壁变为不连续的,壁物质明显降低。对生殖细胞分裂前的位置及精细胞发育中形态变化的意义进行了讨论。     

休眠期和营养期包囊游仆虫的纤毛器骨架及其微管蛋白

应用光镜和透射电镜术 ,显示了包囊游仆虫休眠细胞中纤毛器骨架的形态 ,并对该纤毛虫休眠细胞和营养细胞的纤毛器及其α、β -微管蛋白进行了免疫荧光定位的比较研究。由免疫荧光显微术显示 ,包囊游仆虫形成休眠包囊后 ,背部毛基体完整地按原有模式保存下来 ;纤毛杆解聚后微管蛋白多集中在细胞皮层 ,小部分均匀散布在细胞质中。据所得结果认为 ,包囊游仆虫形成包囊后 ,微管蛋白主要有 3个去向 ,即 :①处于自噬泡内被逐步消化 ;②以“微管蛋白库”的形式分布于细胞皮层及细胞质中 ;③保留在残留的基体中。此外 ,以往研究中发现的棘毛基部纤维网络未被标记上 ,提示这些纤维体系可能不属于微管系统     

体眠期和营养期包裹游仆虫的纤毛器骨架及其微管蛋白

应用光镜和透射电镜术,显示了包囊游仆虫休眠细胞中纤毛器骨架的形态,并对该纤毛虫休眠细胞和营养细胞的纤毛器及其α,β－微管蛋白进行了免疫荧光定位的比较研究。由免疫荧光显微术显示,包囊游仆虫形成休眠包囊后,背部毛基体完整地按原有模式保存下来;纤毛杆解聚后微管蛋白多集中在细胞皮层,小部分均匀散布在细胞质中,据所得结果认为,包囊游仆虫形成包囊后,微管蛋白主要有3个去向,即：91）处于自噬泡内被逐步消失;（2）以“微管蛋白库”的形式分布于细胞皮层及细胞质中;（3）保留在残留的基体中,此外,以往研究中发现的棘毛基部网络未被标记上,提示这些纤维体系可能不属于微管系统。     

Status of THz-to-Visible Nanospectroscopy Development

Quite unexpectedly, THz and infraredspectroscopy has now a real chance to solveproblems in the nanosciences. This rests ona new microscope technique that overcomesthe Abbe diffraction limit, by using thenear field of a metal antenna in closeproximity to a scanned sample surface. HereI briefly summarize present activities inthe microwave, mid-infrared and visiblespectral ranges. It seems straightforwardand highly desirable to fill the existinggap between about 20 GHz and 20 THz, andattain spatial resolution of 10 nm andbelow also in this important part of theelectromagnetic spectrum.     

Fluorescence Microscopy Gets Faster and Clearer: Roles of Photochemistry and Selective Illumination

Significant advances in fluorescence microscopy tend be a balance between two competing qualities wherein improvements in resolution and low light detection are typically accompanied by losses in acquisition rate and signal-to-noise, respectively. These trade-offs are becoming less of a barrier to biomedical research as recent advances in optoelectronic microscopy and developments in fluorophore chemistry have enabled scientists to see beyond the diffraction barrier, image deeper into live specimens, and acquire images at unprecedented speed. Selective plane illumination microscopy has provided significant gains in the spatial and temporal acquisition of fluorescence specimens several mm in thickness. With commercial systems now available, this method promises to expand on recent advances in 2-photon deep-tissue imaging with improved speed and reduced photobleaching compared to laser scanning confocal microscopy. Superresolution microscopes are also available in several modalities and can be coupled with selective plane illumination techniques. The combination of methods to increase resolution, acquisition speed, and depth of collection are now being married to common microscope systems, enabling scientists to make significant advances in live cell and in situ imaging in real time. We show that light sheet microscopy provides significant advantages for imaging live zebrafish embryos compared to laser scanning confocal microscopy.     

Virtual slit scanning microscopy

We present a novel slit scanning confocal microscope with a CCD camera image sensor and a virtual slit aperture for descanning that can be adjusted during post-processing. A very efficient data structure and mathematical criteria for aligning the virtual aperture guarantee the ease of use. We further introduce a method to reduce the anisotropic lateral resolution of slit scanning microscopes. System performance is evaluated against a spinning disk confocal microscope on identical specimens. The virtual slit scanning microscope works as the spinning disk type and outperforms on thick specimens.     

Recent Developments in Correlative Super-Resolution Fluorescence Microscopy and Electron Microscopy

The recently developed correlative super-resolution fluorescence microscopy (SRM) and electron microscopy (EM) is a hybrid technique that simultaneously obtains the spatial locations of specific molecules with SRM and the context of the cellular ultrastructure by EM. Although the combination of SRM and EM remains challenging owing to the incompatibility of samples prepared for these techniques, the increasing research attention on these methods has led to drastic improvements in their performances and resulted in wide applications. Here, we review the development of correlative SRM and EM (sCLEM) with a focus on the correlation of EM with different SRM techniques. We discuss the limitations of the integration of these two microscopy techniques and how these challenges can be addressed to improve the quality of correlative images. Finally, we address possible future improvements and advances in the continued development and wide application of sCLEM approaches.     

Perfectly registered multiphoton and reflectance confocal video rate imaging of in vivo human skin

We present a multimodal in vivo skin imaging instrument that is capable of simultaneously acquiring multiphoton and reflectance confocal images at up to 27 frames per second with 256 × 256 pixel resolution without the use of exogenous contrast agents. A single femtosecond laser excitation source is used for all channels ensuring perfect image registration between the two‐photon fluorescence (TPF), second harmonic generation (SHG), and reflectance confocal microscopy (RCM) images. Images and videos acquired with the system show that the three imaging channels provide complementary information in in vivo human skin measurements. In the epidermis, cell boundaries are clearly seen in the RCM channel, while cytoplasm is better seen in the TPF imaging channel, whereas in the dermis, SHG and TPF channels show collagen bundles and elastin fibers, respectively. The demonstrated fast imaging speed and multimodal imaging capabilities of this MPM/RCM instrument are essential features for future clinical application of this technique. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

A developed temporal focusing‐based multiphoton excitation microscope (TFMPEM) has a digital micromirror device (DMD) which is adopted not only as a blazed grating for light spatial dispersion but also for patterned illumination simultaneously. Herein, the TFMPEM has been extended to implement spatially modulated illumination at structured frequency and orientation to increase the beam coverage at the back‐focal aperture of the objective lens. The axial excitation confinement (AEC) of TFMPEM can be condensed from 3.0 μm to 1.5 μm for a 50 % improvement. By using the TFMPEM with HiLo technique as two structured illuminations at the same spatial frequency but different orientation, reconstructed biotissue images according to the condensed AEC structured illumination are shown obviously superior in contrast and better scattering suppression. Picture : TPEF images of the eosin‐stained mouse cerebellar cortex by conventional TFMPEM (left), and the TFMPEM with HiLo technique as 1.09 μm^?1 spatially modulated illumination at 90° (center) and 0° (right) orientations.

     

Portable optical‐resolution photoacoustic microscopy for volumetric imaging of multiscale organisms

Photoacoustic microscopy (PAM) provides a fundamentally new tool for a broad range of studies of biological structures and functions. However, the use of PAM has been largely limited to small vertebrates due to the large size/weight and the inconvenience of the equipment. Here, we describe a portable optical‐resolution photoacoustic microscopy (pORPAM) system for 3‐dimensional (3D) imaging of small‐to‐large rodents and humans with a high spatiotemporal resolution and a large field of view. We show extensive applications of pORPAM to multiscale animals including mice and rabbits. In addition, we image the 3D vascular networks of human lips, and demonstrate the feasibility of pORPAM to observe the recovery process of oral ulcer and cancer‐associated capillary loops in human oral cavities. This technology is promising for broad biomedical studies from fundamental biology to clinical diseases.      

Microscopy basics and the study of actin–actin-binding protein interactions

Actin is a multifunctional eukaryotic protein with a globular monomer form that polymerizes into a thin, linear microfilament in cells. Through interactions with various actin-binding proteins (ABPs), actin plays an active role in many cellular processes, such as cell motility and structure. Microscopy techniques are powerful tools for determining the role and mechanism of actin–ABP interactions in these processes. In this article, we describe the basic concepts of fluorescent speckle microscopy, total internal reflection fluorescence microscopy, atomic force microscopy, and cryoelectron microscopy and review recent studies that utilize these techniques to visualize the binding of actin with ABPs.