整体透明技术在植物生物学中的应用实例及其剖析

通过采用不同的透明剂和透明方法,对番茄（Lycopercicon esculenturm）侧根原基、加拿大一枝黄花（Solidago canadensis）与一枝黄花（S．decurens）的亲和性识别反应和苏门白酒草（Conyza sumatrensis）的胚珠发育过程进行了观察,提供了整体透明技术在植物生物学中的应用实例,简要回顾了该技术在植物生殖生物学中的应用和发展状况,分析了该技术在植物生物学应用中的优势和不足,探讨了该技术应用中一些具体的技术环节,如透明剂的选择和使用以及与特殊用途显微镜的配合使用等方面的问题,并对该技术的应用前景进行了展望。     

光学透明技术在植物多尺度成像中的应用

光学透明技术是一种通过各种化学试剂,将原本不透明的生物样本实现透明化,并在光学显微镜下深度成像的技术。结合多种光学显微成像新技术,光学透明技术可对整个组织进行成像和三维重建,深度剖析生物体内部空间特征与形成机制。近年来,多种植物光学透明技术和多尺度成像技术被陆续研发,并取得了丰硕的研究成果。该文综述了生物体光学透明技术的基本原理和一些新技术,重点介绍基于光学透明技术开发的新型成像方法及其在植物成像与细胞生物学中的应用,为后续植物整体、组织或器官的透明、成像与三维重构及功能研究提供理论依据和技术支持。     

整体染色透明技术在植物学研究中的应用简介

整体染色透明技术在植物学研究中的应用简介李婷婷（学生）（华南农业大学农学系广州５１０６４２）１整体染色透明技术的历史及发展整体染色透明技术是近２０年发展起来的制片新技术。它不需要切片,只经过透明剂透明或染色透明便可在显微镜下观察,是一种较石蜡制片简便...     

植物生殖生物学研究的一些进展

简介参与花粉与柱头相互识别的s位点编码产物、花药城毡层组织特异启动子的克隆及应用、花粉粒内基因表达、卵细胞高体显微操作和合于形成前后的基因表达五个方面的研究动态和进展。     

植物的生殖讲座（八）——植物生殖生物学研究进展

植物的生殖讲座（八）──植物生殖生物学研究进展陈祖铿（中国科学院植物研究所北京１０００４４）植物生殖生物学是研究植物有性生殖过程的一门分支学科,它涉及性细胞发育、传粉与受精,以及胚胎发生等。从广义讲,可以包括各种形式的营养体无性繁殖和孢子生殖过程。花...     

荧光显微术在当代植物细胞生物学研究中的应用

自从八十年前第一次在显微镜下观察到生物组织经紫外线照射后发射荧光的现象以来,荧光显微术不断获得进步,现已发展成细胞生物学中一个重要的研究手段。高度的灵敏性和专一性、制样与观察程序的简便、尤其是适宜于活细胞研究等特点,是它所具有的独特长处。荧光显微术特别是免疫荧光技术在现代医学生物学研究中的应用是一个     

化石植物生殖生物学研究的现况和前景

植物的生殖结构及其本质,在植物(特别是高等维管植物)的系统演化和分类的研究中,占有特别重要地位,以至常被当作系统分类的主要依据。然而对化石植物而言,生殖结构的发现是相当困难的事,要准确地了解其生殖本质则更困难。不过由于古植物学家们的努力,关于化石植物生殖本质的秘密正被逐渐揭示,工作也     

组织透明技术在植物解剖学和植物发育生物学实验教学中的应用

整体组织透明和组织切片技术是研究植物组织和器官的结构及其发育过程的常用实验手段。该文针对本科生植物学课程的侧根、胚珠(特别是胚囊)以及胚胎发育内容设计了相应的实验教学课程。该实验以拟南芥幼苗、不同发育时期的花蕾和种子为材料,经固定、脱色和组织透明,利用正置显微镜观察侧根、胚珠以及胚胎的发育过程。根组织经透明后,在显微镜下可以清晰地观察到从侧根原基的建成到侧根发生的八个发育阶段。各个发育时期的胚珠经组织透明后,在显微镜下可以清晰地观察到大孢子母细胞、功能性大孢子(FG1期)以及FG2~FG7期的胚囊。各个发育时期的种子经透明后,在显微镜下可以清晰地观察到胚胎发育的1细胞期、8细胞期、球形期、心形期、鱼雷期、拐杖期和成熟期。这些实验结果可以使学生对拟南芥等双子叶植物的侧根、胚珠以及胚胎发育过程有较为系统的了解。该实验方法在其他较小的双子叶植物中也适用。     

植物实验生殖生物学与生殖细胞工程:现在与未来

植物实验生殖生物学是植物实验胚胎学发展的新阶段,其主要特征为操作技术水平的提高与多学科综合性研究的加强。花粉原生质体、生殖细胞、精子,胚囊、卵细胞的操作、雌雄配子体外融合、配子-体细胞杂交等,代表了当前的主要研究趋势。实验生殖生物学与基因工程相结合,开辟了植物生殖工程新技术领域的前景。对生殖工程的意义与内容提出了轮廓设想。     

The Subventricular Zone En-face: Wholemount Staining and Ependymal Flow

The walls of the lateral ventricles contain the largest germinal region in the adult mammalian brain. The subventricular zone (SVZ) in these walls is an extensively studied model system for understanding the behavior of neural stem cells and the regulation of adult neurogenesis. Traditionally, these studies have relied on classical sectioning techniques for histological analysis. Here we present an alternative approach, the wholemount technique, which provides a comprehensive, en-face view of this germinal region. Compared to sections, wholemounts preserve the complete cytoarchitecture and cellular relationships within the SVZ. This approach has recently revealed that the adult neural stem cells, or type B1 cells, are part of a mixed neuroepithelium with differentiated ependymal cells lining the lateral ventricles. In addition, this approach has been used to study the planar polarization of ependymal cells and the cerebrospinal fluid flow they generate in the ventricle. With recent evidence that adult neural stem cells are a heterogeneous population that is regionally specified, the wholemount approach will likely be an essential tool for understanding the organization and parcellation of this stem cell niche.     

Wholemount immunohistochemistry for revealing complex brain topography

The repeated and well-understood cellular architecture of the cerebellum make it an ideal model system for exploring brain topography. Underlying its relatively uniform cytoarchitecture is a complex array of parasagittal domains of gene and protein expression. The molecular compartmentalization of the cerebellum is mirrored by the anatomical and functional organization of afferent fibers. To fully appreciate the complexity of cerebellar organization we previously refined a wholemount staining approach for high throughput analysis of patterning defects in the mouse cerebellum. This protocol describes in detail the reagents, tools, and practical steps that are useful to successfully reveal protein expression patterns in the adult mouse cerebellum by using wholemount immunostaining. The steps highlighted here demonstrate the utility of this method using the expression of zebrinII/aldolaseC as an example of how the fine topography of the brain can be revealed in its native three-dimensional conformation. Also described are adaptations to the protocol that allow for the visualization of protein expression in afferent projections and large cerebella for comparative studies of molecular topography. To illustrate these applications, data from afferent staining of the rat cerebellum are included.     

Optimized Protocol for Retinal Wholemount Preparation for Imaging and Immunohistochemistry

Working with delicate tissue can be a complicating factor when performing immunohistochemical assessment. Here, we present a method that utilizes a ring-supported hydrophilized PTFE membrane to provide structural support to both living and fixed tissue during immunohistochemical processing, which allows for the use of a variety of protocols that would otherwise cause damage to the tissue. First, this is demonstrated with bolus loading of fluorescent markers into living retinal tissue. This method allows for quick visualization of targeted structures, while the membrane support maintains tissue integrity during the injection and allows for easy transfer of the preparation for further imaging or processing.Second, a procedure for antibody staining in tissue fixed with carbodiimide is described. Though paraformaldehyde fixation is more common, carbodiimide fixation provides a superior substrate for the visualization of synaptic proteins. A limitation of carbodiimide is that the resulting fixed tissue is relatively fragile; however, this is overcome with the use of the supporting membrane. Retinal tissue is used to demonstrate these techniques, but they may be applied to any fragile tissue.     

Wholemount histofluorescence of catecholamine-containing neurones in a hemipteran brain

Whole brains from Rhodnius prolixus treated with glyoxylic acid display catecholamine histofluorescence in eleven bilaterally distributed clusters of neurones, and in three circumscribed neuropile regions. This simple method offers resolution of some features not detected in studies on sectioned insect tissue; cephalic neurosecretory cells in chronically starved animals are histofluorescent and diverse catecholamine-handling fibres converge within the brain of this insect.     

Techniques for Clearing Ovules for Studies of Megagametophyte and Early Postfertilization Development in Rhododendron

Using ovule clearing, more than 33,600 ovules of Rhododendron nuttallii T. W. Booth (Ericaceae) were examined for megagametophyte and early postfertilization stages at daily intervals from anthesis until 3 weeks after pollination. Pretreatments with amyloglucosidase to digest integumentary and gametophyte starch and Stockwell's bleach to remove tannins from the integumentary epidermis were necessary. Ovules were cleared by a combination or modifications of Heir's four-and-a-half or Stelly's hem-alum-methyl salicylate techniques and were observed using differential interference contrast optics. The method proved suitable for large-scale quantitative studies of ovule development and fertilization. A general protocol is suggested as a starting point for ovule clearing studies.     

例析遗传学中的判断性实验设计问题

将遗传学知识与实验设计两者结合起来进行考查,是近几年生物学高考命题中的一大看点。该类试题注重严密的逻辑思维、缜密的演绎推理以及开放性的实验设计等能力考查,能较好地反映出考生分析、综合、判断和推理等思维品质,因此该类试题往往成为了考生答题的瓶颈。在备考中,教师要善于进行专题性的归纳总结,多在知识系统化、条理化和思维逻辑化上做研究,切实提高考生辨证思维能力、逻辑推理能力以及驾驭知识的能力。本文就高中遗传学中的几个判断性实验设计问题归类例析如下。