透射电子显微镜(TEM)技术在大孢子化石超微结构研究中的应用

透射电子显微镜(TEM)具有优异的纳米尺度显微成像性能,因此被广泛应用于材料科学、物理学和生物学等学科的超微结构研究领域。文中根据应用TEM技术研究大孢子化石壁超微结构的实践体会,系统归纳和总结了大孢子化石TEM样品制备的前处理过程,主要步骤包括:材料选取、梯度脱水、包埋剂配比、梯度渗透、包埋、聚合、超薄切片和染色。运用上述TEM实验技术,文中以云南省曲靖市沾益区龙华山剖面中泥盆统吉维特阶上双河组分散拟网龙华山大孢(Longhuashanispora reticuloides Lu and Ouyang, 1978)的壁层超微结构为研究案例,揭示了这类孢子壁主要由三层薄壁组成,包括内部基底层、中部疏松层和外部致密层,其射线唇基底层下部具有近平行排列的多细纹带结构。此类大孢子的多细纹带结构与从加拿大新不伦瑞克省下泥盆统埃姆斯阶Campbellton组同孢植物化石Leclercqia complexa Banks et al., 1972中发现的原位小孢子射线唇基底层下部的多细纹带结构特征非常相似。此外,两者都具有相似的完全弓形脊和远极表面刺瘤状二型纹饰。因此综合外壁超微结构和纹饰形态特征...     

新技术,新方法在孢粉学中的应用

最近几年来,扫描电镜(SEM)、透射电镜(TEM)、微机(Computer)和荧光显微镜(Fluorescence microscope)等新技术、新方法,在孢粉学领域中得到广泛应用。 一、扫描电镜孢粉样品的制作方法 扫描电镜与普通光学显微镜比较,它有放大倍数范围大、分辨率高、立体感强,和样品的制备手续简便等特点。所以,从扫描电镜问鼎到现在,虽然才仅仅经历了二十多年的时间,但它却像雨后春笋般地出现在世界各地,在孢粉学中获得了极为广泛的运用,从而使孢粉学这门新兴边缘学科,获得了突飞猛进的发展。     

(木奈)李的孢粉学初步研究

为了解李的起源,应用扫描电镜对7份李(Prunus salicina L.var.cordata Y.He)、2份中国李(P.salicina Lindl.var.salicina)、1份樱桃李(P.cerasifera Ehrhart.)及3份杂种李的花粉形态进行观察和聚类分析,开展李的孢粉学研究。结果表明,这13份李属植物的花粉具条纹-穴状或脑纹-穴状纹饰,花粉形状均为长球形或超长球形,极面观为三裂圆形,具3条萌发沟,萌发沟以等间距环状分布,属N3P4C5类型。李的P/E(1.82~2.02)、花粉大小(823.70~970.42μm2)介于供试的其他6份李属植物之间,但其穿孔密度(0.520~1.141μm–2)除早圆油和早熟油外均远大于其他6份李材料;李花粉外壁具条脊,表面均形成大小、疏密不等的穿孔,且在条脊隆起、脊沟深窄等方面表现出了丰富的多样性,其中油早熟品系和晚熟品系的花粉纹饰、穿孔密度等还表现出了2种特征,因此,李的孢粉学特征可作为分类鉴定的一个重要依据。在遗传距离6.19水平上,13份李属植物可以分为3个类群,每个类群中均有李分布,李与李属间具有较近的亲缘关系。综合李花粉形态特征和聚类分析结果,推测李具起源于李属杂交后代的可能。     

中国孢粉学发展的契机与抉择(兼论文集后记)

回顾中国孢粉学的发展历程,认为孢粉学在中国的兴起和发展是与国家的经济建设,以及改革开放政策和知识创新工程的先后实施所带来的契机紧密相联系.呼吁中国孢粉工作者能正面抉择,不断创新,共同促成学科的理论发展和学科间的联合.简要介绍论文集内容,除本文外,含回顾性文章2篇、地层孢粉论文8篇、表土花粉的陆地植被覆盖度模拟和孢粉通量论文各1篇、现代花粉形态论文3篇.     

植物透射电镜样品制备技术探讨

针对植物细胞壁组织坚硬,透射电镜样品的制备难度大的问题,在样品制备过程中逐步改进了常规技术：其中通过延长锇酸固定时间,提高了植物微细结构保存效果;利用减少树脂渗透浓度的方法,改善了树脂包埋头的切割性能.避免了透射电镜观察时,微细结构不清晰和切片破损现象,提高了样品分辨率和观察范围.试验结果表明,此方法能节约透射电镜样品的制备成本、缩短样品制备周期和提高样品制备质量.     

电子显微镜虚拟仿真实验的建设与应用

电子显微镜是利用电磁透镜聚焦成像的一种具有超高分辨率的大型电子光学仪器。但由于电子显微镜的结构精密、价格昂贵、运行和维修的成本较高等特点,使其在本科实验教学与实践培训中开展困难且存在着实验资源欠缺、教学成本高、仪器内部构造不易观察、实验时间过长等问题。为了解决这一难题,依据电子显微镜的成像原理,借鉴和参考最新的科研成果和技术应用案例,设计了教学目标并构建了知识体系,采用新一代信息技术建设了电子显微镜虚拟仿真实验项目。该实验包括电子显微镜超薄切片技术、负染色技术和线上答题等模块,可以实现实验内容的在线操作、自主学习、在线考核并对学生学习效果的全程追踪和综合性评价等功能。虚实结合的教学实践表明,该项目有效拓展了学生的学习空间与知识体系,丰富了实验教学与实践培训的内容和手段,形成了以学生为中心的自主学习教学资源,显著提高了实验教学效果,有利于基础学科创新拔尖人才的培养。     

29个蔷薇属植物的孢粉学研究

采用扫描电镜技术对蔷薇属25个种及4个月季品种的花粉形态进行观察分析,探讨单叶亚属及蔷薇亚属中的芹叶组、合柱组、桂味组、月季组及月季品种之间的亲缘关系。研究表明:(1)所有被测样品的花粉都为具三孔沟的N3P4C5型单粒花粉,形状包括近球形、长球形及超长球形,其中大部分都为极轴较长的超长球形,以小檗叶蔷薇(P/E=2.29)最长。(2)在疏花蔷薇、托木尔蔷薇以及重瓣黄刺玫中发现有花粉二型现象。(3)被测样品外壁纹饰主要包括条纹、孔穴-条纹及近孔网纹3类,条纹类型主要出现在芹叶组中,而孔穴-条纹类型则为蔷薇属植物较普遍的纹饰。(4)聚类分析表明,小檗叶蔷薇与其它种亲缘较远,被测月季品种与月季组关系较近,芹叶组与合柱组关系较近,桂味组中的弯刺蔷薇与其他蔷薇的纹饰有明显区别,被单独分开。     

成都地区19个木芙蓉品种的孢粉学研究及其分类学意义

为了解成都地区木芙蓉(Hibiscus mutabilis)品种间的亲缘关系,采用扫描电镜观察了19个木芙蓉品种的花粉显微结构,并探讨其分类学意义.结果表明,19个木芙蓉品种的花粉均为大粒,主要为圆球形,具散孔,外壁纹饰均为刺状纹饰,表面具有颗粒状突起.木芙蓉品种间在花粉粒大小、外壁纹饰上具有一定差异,可以作为品种分类...     

基于文献计量学的近30年(1990-2020)第四纪孢粉学研究现状与展望

第四纪孢粉学利用现代和化石孢粉重建第四纪古植被、古气候和古环境,属于可持续发展的前沿学科领域。本文基于CiteSpace软件数据可视化和计量分析模块,以CNKI (中国知网)数据库与Web of Science核心数据库中1990–2020年间的共计6081篇第四纪孢粉学相关文献为基础,梳理了近30年来第四纪孢粉学的过去和现在,并探讨了未来发展趋势。研究结果表明,近30年第四纪孢粉学领域发文量呈增加趋势,中国的总发文量和国际影响力整体呈增长趋势;第四纪孢粉学研究主要集中于各大高校和科研院所,研究成果有明显的聚集性;中国科学院是外文发文量最多的国际机构。第四纪孢粉学近30年的研究主要涉及古植物学、古地理学、古生态学和古气候学等领域,古植被、古气候、古环境的定性和定量重建一直是国内外学者关注和研究的重点。表土花粉研究、花粉与人类活动、亚洲季风演变和孢粉分类学是近几年发刊的热点。虽然一些关键词如花粉产量、代表性以及现代花粉与植被和气候的定量关系等出现频次低,但对第四纪孢粉学十分重要。未来还需要进一步加强湖泊表层沉积物的花粉研究,探讨花粉与植被、气候的定量关系,为准确解译化石花粉提供坚实基础。...     

皱纹盘鲍受精过程的电镜观察

本文用透射电镜观察了皱纹盘鲍的受精过程。鲍卵子的胶膜使精子活化,并诱发了顶体反应,卵黄膜使顶体反应达到高潮。精子入卵后,卵发生皮层反应并形成受精膜开 减数分裂。此外,还观察到鲍的多精入卵现象。     

ELECTRON MICROSCOPE STUDY OF MITOSIS IN SEA URCHIN BLASTOMERES

The fine structure of cells at different stages of the mitotic cycle was studied in the blastomeres of 6-hour-old embryos of the sea urchin Strongylocentrotus purpuratus. The material was fixed in 1 per cent osmium tetroxide in sea water, buffered with veronal-acetate to pH 7.5, embedded in Araldite, and sectioned with glass knives. The aster, as it forms around the centriole, has the appearance of the endoplastic reticulum, with elements oriented radially from the centrosphere to the periphery of the cell. Anaphase structures described include the kinetochores, with bundles of fine filaments extending toward the centrioles, as well as continuous filaments passing between the chromosomes. Two cylindrical centrioles composed of parallel rods are present in each of the anaphase asters. At late anaphase, elements of the endoplasmic reticulum condense on the surface of the chromosomes to form a double membrane which already at this stage possesses pores or annuli. At telophase bundles of continuous filaments can be seen in the interzonal region. These filaments, as well as those associated with the chromosomes, have a diameter of approximately 15 mµ, and appear physically different from the astral structure.     

AN ELECTRON MICROSCOPE STUDY OF LYMPHATIC TISSUE IN RUNT DISEASE

The thymus, spleen, and lymph nodes were studied in runt disease induced by a graft of intravenously injected homologous splenic cells into newborn rats and mice. Adult Long-Evans cells (70 x 10⁶) were injected into Sprague-Dawley rats. Adult DBA cells (7 x 10⁶) were injected into C57BL/6 mice. Runted rats were sacrificed at 14 to 28 days of age; mice at 10 to 20 days. The thymic cortex is depleted of small lymphocytes. Those remaining are severely damaged and phagocytized. Evidence of damage includes swelling of mitochondria, myelin figure formation, margination of chromatin, and sharp angulation in nuclear contour. Large numbers of macrophages are present. Epithelial-reticular cells which envelop small cortical blood vessels are often retracted, with the result that the most peripheral layer in the thymic-blood barrier suffers abnormally large gaps. Lymphocytes of the periarterial lymphatic sheaths of spleen and of the cortex of lymph nodes are reduced in number and damaged. Vast numbers of plasma cells and many lymphocytes are evident throughout lymph nodes, in the periarterial lymphatic sheaths, and in the marginal zone and red pulp of the spleen. Plasma cells are of different sizes, the larger having dilated sacs of endoplasmic reticulum. Lymphocytes are small to medium in size. They contain, in varying quantity, ribosomes and smooth membrane-bounded cytoplasmic vesicles approximately 350 to 500 A in diameter. Most plasma cells and lymphocytes are damaged and many of these are phagocytized. Many lymphocytes in lymph nodes, however, show no evidence of damage. Reticular cells and other fixed cells of the connective tissues seldom appear affected. Thus, the major cell types reacting in runt disease are lymphocytes, plasma cells, and histiocytes or macrophages. It appears, therefore, that both the delayed and immediate types of sensitivity play a part in this disease.     

体外神经干细胞克隆球的超微结构-透射电镜观察

为观察培养的神经干细胞克隆球内部的超微结构特征,采用无血清培养技术,在体外进行小鼠纹状体神经干细胞克隆球的培养传代,经过免疫细胞化学鉴定后,对单一的神经干细胞克隆球进行固定,常规透射电镜观察。结果表明,神经干细胞可以在bFGF等生长因子存在的情况下,在无血清培养液内增殖生成悬浮状态的神经干细胞克隆球,这种克隆可被诱导生成神经细胞和神经胶质细胞,电镜下,神经干细胞克隆球内部细胞相互间可形成特化的膜性结构,细胞内可有小泡出现,部分细胞有凋亡的形态。     

中华绒螯蟹神经细胞和胶质细胞的光镜及电镜观察

中华绒螯蟹神经节存在神经细胞和胶质细胞两类细胞。神经细胞膜为３层,细胞质由内质网和核蛋白体的聚合体的所构成的尼氏体,高尔基体,线粒体,液泡等细胞器组成,核膜双层,核仁明显,胶质细胞内含有似神经细胞的细胞器结构,根据细胞核的形态及核内染色质的分布情况,可把胶质细胞分成３类：星形胶质细胞核最大,卵圆形,少突胶质细胞核较小,圆形;小胶质细胞核最小,呈三角形或卵圆形,神经细胞的数量与附肢的生理活动强弱有一     

AN ELECTRON MICROSCOPE STUDY OF RADIATION DAMAGE IN THE MOUSE OOCYTE

Cytological changes occurring in young oocytes of the mouse, following whole-body x-irradiation, have been examined by light and electron microscopy. Two minutes after a dose of 200 r, a drop occurs in the number of mitochondria per oocyte. The normal number of mitochondria is restored in the next few minutes. Five to 6 hours after a dose of 7 r, and 30 to 45 minutes following a dose of 200 r, all the oocytes are markedly contracted. In the next 2 hours (at a dose of 7 r), some cells shrink further and become pycnotic, while others expand and show signs of karyolysis. Of the expanded cells, some (about 50 per cent of total young oocytes) become morphologically normal. In both pycnotic and karyolytic nuclei, the nucleolus was contracted but without loss of its fibrillar structure. The contracted nucleolus appeared similar to those seen in some nearly mature normal oocytes (these oocytes have a high natural death rate). No other cell type in the ovary was affected by doses of x-rays up to 200 r. Observations on the nucleus of the normal oocyte are included. In the dictyate stage of meiotic prophase the chromosomes were dispersed into bundles of 100 A microfibrils. The main component of the nucleolus was found to be tightly coiled fibrils of 60 to 100 A, which appear to have a close relationship with the microfibrils of the chromosomes.     

AN ELECTRON MICROSCOPE STUDY OF SILVER NITRATE REDUCTION IN LEAF CELLS

Brown , W. V., H. Mollenhauer , and C. Johnson . (U. Texas, Austin.) An electron microscope study of silver nitrate reduction in leaf cells. Amer. Jour. Bot. 49(1): 57–63. Illus. 1962.—As reported earlier in many studies, AgNO₃ is reduced quickly by the living chloroplasts of angiosperms. Electron microscope study has resolved the conflict of opinions concerning the exact location of the silver particles. Reduction of AgNO₃, as indicated by location of silver particles, occurs within the chloroplasts but not within the grana or pure stroma; it appears to be associated with the intergranal (also called stroma) lamellae. Silver particles are formed also at the surfaces of the cell wall, both in the middle lamella and at the inner surface, and also within plasmodesmata. It is concluded that chlorophyll is probably not involved directly in the reduction. There is some slight support for the popular hypothesis that ascorbic acid may be the chief reducing agent.