化石植物研究方法简介

古植物学是研究古代植物和它们生活的科学。重现植物界历史的方法之一是研究岩层中的化石植物。植物以各种形式保存在地壳之中,在保存期间发生了各种物理、化学变化。不同的环境和沉积过程也会导致化石出现种种不同的保存形式。针对不同的类型,必须采用不同的技术处理,以便从中提取信息,找出同一植物各分散器官之间的联系,从整体进行研究和重建。１　角质层的处理方法化石角质层为地质史上真正生活过的植物活体上的原物,它与植物生活的环境有一定的联系,可作为探讨属种间的自然亲缘关系的依据。化石角质层有几种处理方法（叶美娜,１…     

四种植物花蜜腺的解剖学研究

通过对石竹、庐山小檗、活血丹和紫云英四种植物花蜜腺的外部形态、显微切片和扫描电镜观察,了解到石竹花蜜腺是雄蕊类型,蜜汁通过角质层间小孔分泌;庐山小檗花蜜腺是花被类型,其每个分泌表皮细胞的外切向壁中央向内凹陷与角质层之间形成角质层下空隙,蜜汁由表皮细胞分泌后贮存于角质层下空隙中,在蜜汁积累过程中不断增加对角质层的压力,最后冲破角质层分泌到体外;活血丹和紫云英花蜜腺属于花托类型,前者蜜汁从分泌表皮细胞通过角质层直接渗出,后者蜜汁通过变态气孔分泌。本试验的四种植物花蜜腺中维管组织有三种不同类型。     

景深合成技术在植物光学微形态研究上的应用

介绍了利用电脑景深合成技术在光学显微镜上获得植物标本大景深图片的摄影方法。概述了此方法获取植物光学微形态特征清晰图片的基本步骤和优缺点。该方法制片简单,可利用最普通的生物显微镜、电子目镜结合计算机进行摄影,成本低,观察特征明显,获得的图片具有真色彩,并可拍出植物最鲜活的状态。此方法有可能成为未来植物微形态研究的一个重要手段。     

三维反卷积显微成像技术浅谈

三维宽场反应卷积显微成像技术是应用光学切片方法获取三维标本的二维图像序列,然后通过反卷积图像处理方法进行图像恢复,进而进行三维重建的一种以光学技术和图像处理技术为核心的显微成像方法。本文讲述了光学切片的基本原理,给出了反卷积处理中点扩展函数的理论模型和实验测试方法,然后对现存的反卷积算法做了对比。最后,文章对这一领域的发展趋势作了预测。     

水杉属和红杉属化石叶表皮鉴定参照系的特殊性

杉科植物的许多属种在小枝的形态和叶片排列上相似,而杉科植物的化石标本多保存为枝叶形式。表皮的特征作为压型化石枝叶标本细胞信息的重要来源,甚至是惟一来源。一直作为杉科植物化石分类鉴定的主要依据。水杉和北美红杉分别是水杉属和红杉属植物化石的惟一现存最近亲缘种,以往关于北美红杉的气孔分布和排列等方面的报道存在分歧,根据作者的研究,北美红杉的表皮特征变异幅度非常广。水杉的气孔分布也与以往报道有差异。利用表皮的特征鉴定杉科植物化石时;不同的处理方法和处理时间,角质层的完整程度和观察数量等均可以影响植物表皮特征的正确获取。     

石蜡切片中同时显示植物细胞内淀粉,蛋白质和脂肪的方法

利用植物显微化学方法,在石蜡切片中同时显示出植物细胞内的淀粉、蛋白质和脂肪,在光学显微镜下可同时观察到这些组分的形态结构和分布。     

基于谱域光学相干层析系统中光谱涨落的指纹获取

指纹因其特异性和稳定性等特点而被称为"证据之首",在案件侦破中起着极其重要的作用。多种物理学、化学和光学技术都可以用于获取现场遗留的指纹,然而这些方法存在一些缺点,如会对指纹造成破坏、潜在的毒副作用、在现场留有痕迹等。利用谱域光学相干层析(spectral domain optical coherence tomography,简记为SD-OCT)技术进行指纹探测具有非接触、对指纹无损伤和高灵敏度的优势,利用OCT系统的相位敏感性我们可以在低对比条件下再现遗留在物体表面的指纹,但处理结果受指纹所在表面高低起伏影响,使得指纹信息对比度降低,难以被分离和提取。本文提出了一种基于干涉光谱涨落的指纹获取方法,只需对OCT系统得到的干涉光谱的涨落进行处理分析,即可得到遗留在物体表面的指纹图案,不需进行相位和表面轮廓高度的求取,算法简单、处理速度快,处理结果不受样品表面高低起伏的影响。实验结果表明,在起伏表面上,使用该方法也能较好地显现遗留在物体表面的指纹图案。     

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。     

中国兜被兰属植物叶表皮微形态特征的研究

利用光学显微镜和扫描电子显微镜观察了中国兜被兰属１２种植物叶表皮的微形态特征,结果表明：该属植物成熟叶片的上下表皮均具角质层,其表面纹饰有直线形、浅波形、深波形和网状４种类型;有的种表皮细胞垂周壁明显凸起;气孔不具副卫细胞,仅分布在叶片下表面,气孔的外拱盖单层,表面光滑。这些特征在属级水平上较为一致,但也表现出种间差异,可以作为种间分类的辅助特征     

三维反卷积显微成像技术浅谈

三维宽场反卷积显微成像技术是应用光学切片方法获取三维标本的二维图像序列,然后通过反卷积图像处理方法进行图像恢复,进而进行三纺重建的一种以光学技术和图像处理技术为核心的业微成橡方法。本讲述了光学切片的基本原理,给出了反卷积处理中点扩展函数的理论模型和实验测试方法,然后对现存的反卷积算法做了对比。对这一领域的发展趋势做了预测。     

Cytological Staining Procedures Applicable to Methacrylate-Embedded Tissues

Experiments indicate that osmic-fixed, plastic-embedded sections are suitable for examination in the light microscope. Nuclei, mitochondia, cellular membranes and cytoplasmic granules are readily demonstrable by phase microscopy. Connective tissue stains permit the identification of elastic and collagenous fibers. Glycogen and other carbohydrate-containing structures are demonstrable by the periodic acid-Schiff and the ammoniacal silver nitrate procedures. It is, therefore, possible to cross-check individual structures by comparing alternate thick and thin sections, examined in the light microscope and electron microscope respectively. Several other advantages pertain to plastic embedded tissues. The sections compare favorably in translucency and in their lack of distortion with material embedded in celloidin, yet the procedure is simpler and much more rapid. Sections of any desired thinness can be prepared, and alternate thick and thin sections are easily forthcoming. When examined in the phase-contrast microscope, mitochondrial preparations become routinely available without the uncertainties of most of the mitochondrial staining methods. It appears, therefore, that plastic embedding should find a useful place among the methods for light microscopy as well as in the armamentarium of the electron microscopist.     

Preparation of Fossil Bone Specimens for Scanning Electron Microscopy

A method is described for preparing fossil bone specimens for scanning electron microscopy. To obtain bone surfaces suitable for study, material was embedded in Epon 812 and selected faces exposed by grinding were subjected to controlled etching with a 4:1 mixture of 5% HNO₃ and 1% OsO₄, Surfaces thus prepared were further processed by the so-called clearing replicas technique. As a result of this procedure the bone surfaces revealed a network of anastomosing vascular canals the inner surface of whose walls could be examined in the scanning electron microscope. By etching extremely thin ground sections of bone stuck to plastic tape the contents of vascular canals as well as osteocytes can be isolated. This method ensures the good preservation of spatial relations between bone elements essential for studies of fossil bones, which an sometimes very brittle.     

Surface Printing of Plant Leaves for Phylogenetic Studies

A solution of plastic consisting of toluene, 720 ml; methanol, 180 ml; ethyl cellulose (Ethocel, standard 7 CPS), 250 gm; and Dow resin 276 V-2, 75 gm is applied to a leaf surface which has been dampened with toluene. The plastic is spread to a thin film with the edge of a card and allowed to dry. After drying, the plastic may be peeled from the leaf surface and either stored dry in a small envelope or mounted permanently on a microscope slide. Permanent mounts are prepared by placing a small section of the peel from the upper and lower surfaces of the leaf on a microscope slide and covering with a No. 1 cover glass. A small spot of balsam on each corner of the cover glass secures the glass in position. This air mount has proved to be superior to water, glycerol or balsam mounts. Fresh leaves are washed with a mild detergent before application of the plastic. Herbarium specimens are soaked in water overnight to restore the leaf to a semiturgid condition. Five species of different plant families have been illustrated to show the diagnostic features of the surface of the cuticle. An isolated layer of epidermal cells obtained by chemical maceration permitted cell and imprint comparison. The remarkable amount of detail shown by the prints is an aid for phylogenetic studies and may make the recognition of fossil cuticles possible.     

Erratum

A solution of plastic consisting of toluene, 720 ml; methanol, 180 ml; ethyl cellulose (Ethocel, standard 7 CPS), 250 gm; and Dow resin 276 V-2, 75 gm is applied to a leaf surface which has been dampened with toluene. The plastic is spread to a thin film with the edge of a card and allowed to dry. After drying, the plastic may be peeled from the leaf surface and either stored dry in a small envelope or mounted permanently on a microscope slide. Permanent mounts are prepared by placing a small section of the peel from the upper and lower surfaces of the leaf on a microscope slide and covering with a No. 1 cover glass. A small spot of balsam on each corner of the cover glass secures the glass in position. This air mount has proved to be superior to water, glycerol or balsam mounts. Fresh leaves are washed with a mild detergent before application of the plastic. Herbarium specimens are soaked in water overnight to restore the leaf to a semiturgid condition. Five species of different plant families have been illustrated to show the diagnostic features of the surface of the cuticle. An isolated layer of epidermal cells obtained by chemical maceration permitted cell and imprint comparison. The remarkable amount of detail shown by the prints is an aid for phylogenetic studies and may make the recognition of fossil cuticles possible.     

Trilobite cuticle thickness in relation to palaeoenvironment

100 thickness measurements from thin sections of cephala or pygidia of early Ordovician trilobites occurring across an onshore to offshore environmental gradient show that progressively greater maximum cuticle thickness was characteristic of increasingly inshore sites. There is a 40-fold difference between the thinnest and thickest cuticles, and exclusively thin cuticles are confined to the offshore Olenid Biofacies. Variability in cuticle thickness increases offshore to onshore. Environmental control is shown to be more influential on cuticle thickness than is the overall length of the trilobite: some comparatively large trilobites having thin cuticles and small trilobites thick cuticles. The environmental factors which might be responsible for the pattern are briefly discussed. The thin cuticles dominating the offshore Olenid Biofacies were probably appropriate for dysaerobic conditions. Thick cuticles in the most inshore biofacies may have offered protection against predators and turbulence, but the additional presence there of trilobites with thinner cuticles is considered to reflect the greater heterogeneity of the epeiric habitat.     

A rapid procedure for routine double staining of cartilage and bone in fetal and adult animals

A simple, rapid procedure for dual staining of cartilage and bone in rodents, particularly in late gestation, has been developed for routine use. The procedure involves rapid, complete skinning of fresh eviscerated specimens following a 30 sec immersion in a 70 C water bath. The unfixed specimen is stained in a mixture of 0.14% Alcian blue and 0.12% alizarin red S in ethanol and glacial acetic acid. Specimens are then macerated in 2% KOH, cleared and hardened in 1:1 glycerin and distilled water, and stored in pure glycerin. Rapid staining of cartilage only is done in a mixture of 0.08% Alcian blue, glacial acetic acid, and ethanol, with subsequent maceration, clearing, and hardening as in the double staining procedure. Rapid staining of bone only, concurrent with maceration of soft tissue, can be done by placing fresh, unskinned specimens in a diluted mixture of alizarin red S in 2% KOH, with subsequent clearing and hardening in 1:1 distilled water and glycerin. Good quality fetal specimens can be prepared for examination by any of these procedures in a minimum of 11/2-2 days as compared to a minimum of 4-5 days for other procedures. Double stained specimens can be examined for abnormalities of the cartilage as well as bone.     

Estimating sexual dimorphism by method-of-moments

Estimating the degree of sexual dimorphism is difficult in fossil species because most specimens lack indicators of sex. We present a procedure that estimates sexual dimorphism in samples of unknown sex using method-of-moments. We assume that the distribution of a metric trait is composed of two underlying normal distributions, one for males and one for females. We use three moments around the mean of the combined-sex distribution to estimate the means and the common standard deviation of the two underlying distributions. This procedure has advantages over previous methods: it is relatively simple to use, specimens need not be assigned to sex a priori, no reference to living species analogs is required, and the method provides conservative estimates of dimorphism under a variety of conditions. The method performs best when the male and female distributions overlap minimally but also works well when overlap is substantial. Simulations indicate that this relatively simple method is more accurate and reliable than previous methods for estimating dimorphism. © 1996 Wiley-Liss, Inc.     

贵州寒武系第二统杷榔组有壳变形虫的新发现*

有壳变形虫(testate amoebae)的演化历史最早可追溯至新元古代早期, 以该时期北美、华北、挪威以及澳洲等多个地区浅海相碳酸盐岩、页岩中发现的瓶状微体化石(vase-shaped microfossils)为标志。此前认为, 显生宙的有壳变形虫最早出现在早泥盆世。长期以来, 早古生代的地层中未发现这类原生生物的明确化石证据。本研究通过对岩石样品进行常规孢粉酸泡分析处理和切磨岩石薄片, 获取原位保存的化石标本的技术方法, 从贵州东部剑河县交榜剖面出露的寒武系杷榔组(第2统第4阶)中获得数枚有壳变形虫(testate amoebae)化石标本。基于标本的显微形态特征, 并结合激光拉曼光谱等研究, 对原先记述为疑源类的Plagasphaera balangensis和P. sp. A两形态种进行重新认识和描述。由于它们在结构和形态上与一些现生的鳞壳虫目(Euglyphida)有壳变形虫极为相似, 因此将先前定为疑源类的Plagasphaera balangensis和P. sp. A两形态属、种名, 分别修订为Palaeoassulina balangensis gen. et sp. nov.和?Palaeoassulina sp. A。该发现不仅将显生宙有壳变形虫的原有化石记录从晚古生代泥盆纪向前延伸至寒武纪早期, 还为调查研究有壳变形虫的系统演化提供关键的生物化石证据。     

Epidermal and Cuticular Mounts of Plant Material Obtained by Maceration

Mounts of leaf and stem epidermises or bare cuticles, useful in both general anatomical and specialized phylogenetic studies, can be prepared by a maceration process using Jeffrey's solution (equal volumes of 10% aqueous CrO₃ and 10% HNO₃). Leaves, including those of conifers, and stems with cuticles thick enough to maintain integrity when isolated are amenable to this process. Dried specimens are hydrated by boiling in water; fluid-preserved specimens are washed thoroughly in water; fresh specimens need no pretreatment. Specimens are cut to a convenient mount size and trimmed so as to allow adequate and even penetration of the macerating fluid. Laminar leaf segments are left with one edge untrimmed so that upper and lower epidermises remain contiguous. Cylindrical leaf and stem segments are slit lengthwise through about half their thickness. Specimens are macerated in Jeffrey's solution for 1 to 4 days until unwanted tissues are loosened and easily freed from the epidermis (or bare cuticle, if that is desired). The macerating fluid is then washed out completely with changes of water and specimens are stained in a 0.5% aqueous solution of safranin. Dehydration is accomplished with several changes of tertiary butanol. All unwanted tissues not removed by agitation during previous steps of the process are removed by teasing prior to mounting. Specimens are then mounted on slides in Canada balsam and dried in the usual manner.