Determination of the Covariance of Random Sets from Thin Sections

The problem of stereological determination of the covariance of a random set from thin sections is considered. Using results of numerical experiments for two examples, a simple approximation formula is suggested.     

On the Determination of Numerical Density of Cell Organelles Using Mitochondria as an Example

Methods of determining the numerical density of cell organelles described in literature were critically reviewed in a morphometrical and stereological study of muscle cell mitochondria (heart muscle cells, diaphragm cells, sceletal muscle cells). A review of the method described by WEIBEL and GOMEZ showed that the numerical density of the mitochondria depends to a great extent on their shape and not so much on their size distribution. For this reason serial sections should be used to determine the shape factor in biological objects of unknown geometric shape. Generally, the numerical densities of mitochondria determined by using the method proposed by DEHOFF and RHINES were higher than those obtained with the method described by WEIBEL and GOMEZ. This is attributed to certain corrections used in the former method. Elaborate computations are generally involved and the geometric shapes of the object examined must be known in order to determine the numerical density of cell organelles or of other biological structures. The numerous sources of error involved in these methods give this parameter the character of an objective estimate. For this reason it is recommended that the value obtained should be checked by determining a two-dimensional parameter. Our examinations of heart muscle mitochondria showed good agreement between the two parameters.     

Thin Histological Sections Prepared from Large Thick Sections: a New Technique

A method is described for obtaining thin (1 μm) sections for light microscopy from large area thick (100 μm) sections of low viscosity nitrocellulose embedded specimens of human spinal osteoligamentous material.     

The Sizes of Spheres from Profiles in a Thin Slice I. Opaque Spheres

Thin slices through specimens are made into slides for use in microscopy. If a specimen consists of opaque spherical particles in a transparent medium, there will be seen through the slice circular profiles of particles and sections through particles. For a random slice, the size distribution of these profiles can be related to the size distribution of the population of spheres. The extensive literature dealing with this relationship is surveyed. An important generalization of practical importance is made with the introduction of a resolution interval which excludes from observation those profiles that are too large or too small. How this affects the relationship between the profile and sphere size distributions is considered, and some special cases are used as illustrations. In Part II (COLEMAN , 1983) the corresponding results for the case of transparent spheres in an opaque specimen are given.     

Thin Frozen Film Method for Visualization of Storage Proteins in Mature Rice Grains

There are technical difficulties in obtaining intact sections of cereal grains in which mature cells and their subcellular structures are well preserved. Here we describe a simple method for sectioning hard mature rice grains. It makes possible accurate localization of storage proteins in high-quality histological sections of rice endosperm.     

On Kernel Methods of Estimating Marginal Radial and Angular Probability Density Functions

Spatial point patterns which possess a natural origin are considered. Two ways of estimating the marginal radial and angular probability density functions associated with the stochastic process underlying such a pattern are presented. These methods are based on one and two-dimensional kernel functions respectively. The angular density estimate can be used to detect angular trend and to test for angular uniformity within a particular sector about the origin. The two methods of estimation produce essentially the same results. That based on the one-dimensional kernel is recommended because it is computationally simpler.     

Reduced Spectral Density Mapping of a Partially Folded Fragment of E. coli Thioredoxin

Abstract

The backbone dynamics of a partially folded, N-terminal fragment of E. coli thioredoxin were investigated using nuclear magnetic resonance spectroscopy (NMR). Relaxation data were collected at three temperatures and analyzed using reduced spectral density mapping. As temperature was increased, the values for the viscosity normalized J(0) and for J(ω_H) increased, while J(ω_N) decreased. The global trend observed for the viscosity normalized J(0) was consistent with an increase in the hydrodynamic volume of the fragment and suggested the presence of correlated rotational motion in the absence of long range interactions. In addition, the residue specific variation observed for the viscosity normalized J(0) suggested contributions to J(ω) from a range of correlation times that are close to the global correlation time.     

Estimating Stem Volume Using QuickBird Imagery and Allometric Relationships for Open Populus xiaohei Plantations

There has been a great deal of interest in studying the crown of trees using remote sensing data.In this study,crownwidth was extracted from high-resolution QuickBird images for open Populus xiaohei plantations.Regression modelsfor predicting the individual stem volumes of Populus xiaohei were established using extracted crown width,as well asestimated tree parameters(i.e.diameter at breast height[DBH]and tree height)as predictors.Our results indicated thatcrown width could be accurately extracted from QuickBird images using a multi-scale segmentation approach with a meanrelative error of 5.74%,especially for wide-spacing stands.Using either extracted crown width alone or with estimatedDBH and tree height can successfully estimate individual stem volume of Populus xiaohei with the R~2 value ranging from0.87 to 0.92 depending on different model forms.In particular,the two second-order polynomial models(model2 andmodel 6),based on QuickBird image-derived crown widths and estimated DBH and tree heights,respectively,were the bestat describing the relationship between stem volume and tree characteristics.     

Brief Report: Estimating the size and number of autophagic bodies by electron microscopy

Much recent and ongoing research is focused on understanding the mechanisms and regulation of autophagy, a cellular self-degradation pathway with many links to human health. Although many assays exist to measure the total magnitude of autophagy, electron microscopy remains the tool of choice for the determination of the size and the number of autophagosomes formed in a given mutant or under given induction conditions. Here we present a detailed protocol for measuring autophagic bodies in the yeast Saccharomyces cerevisiae by electron microscopy. Furthermore, we present an improved mathematical method for estimating body size and a new method for estimating body number. Finally, we include a discussion of the merits and limitations of these methods and an example of their application to autophagic bodies formed in the ume6∆ strain.     

Preparation of Adult Drosophila Eyes for Thin Sectioning and Microscopic Analysis

Drosophila has long been used as model system to study development, mainly due to the ease with which it is genetically tractable. Over the years, a plethora of mutant strains and technical tricks have been developed to allow sophisticated questions to be asked and answered in a reasonable amount of time. Fundamental insight into the interplay of components of all known major signaling pathways has been obtained in forward and reverse genetic Drosophila studies. The fly eye has proven to be exceptionally well suited for mutational analysis, since, under laboratory conditions, flies can survive without functional eyes. Furthermore, the surface of the insect eye is composed of some 800 individual unit eyes (facets or ommatidia) that form a regular, smooth surface when looked at under a dissecting microscope. Thus, it is easy to see whether a mutation might affect eye development or growth by externally looking for the loss of the smooth surface (''rough eye'' phenotype; Fig. 1) or overall eye size, respectively (for examples of screens based on external eye morphology see e.g.¹). Subsequent detailed analyses of eye phenotypes require fixation, plastic embedding and thin-sectioning of adult eyes.The Drosophila eye develops from the so-called eye imaginal disc, a bag of epithelial cells that proliferate and differentiate during larval and pupal stages (for review see e.g. ²). Each ommatidium consists of 20 cells, including eight photoreceptors (PR or R-cells; Fig. 2), four lens-secreting cone cells, pigment cells (''hexagon'' around R-cell cluster) and a bristle. The photoreceptors of each ommatidium, most easily identified by their light sensitive organelles, the rhabdomeres, are organized in a trapezoid made up of the six "outer" (R1-6) and two "inner" photoreceptors (R7/8; R8 [Fig. 2] is underneath R7 and thus only seen in sections from deeper areas of the eye). The trapezoid of each facet is precisely aligned with those of its neighbors and the overall anteroposterior and dorsoventral axes of the eye (Fig. 3A). In particular, the ommatidia of the dorsal and ventral (black and red arrows, respectively) halves of the eye are mirror images of each other and correspond to two chiral forms established during planar cell polarity signaling (for review see e.g. ³).The method to generate semi-thin eye sections (such as those presented in Fig. 3) described here is slightly modified from the one originally described by Tomlinson and Ready⁴. It allows the morphological analysis of all cells except for the transparent cone cells. In addition, the pigment of R-cells (blue arrowheads in Fig. 2 and 3) can be used as a cell-autonomous marker for the genotype of a R-cell, thus genetic requirements of genes in a subset of R-cells can readily be determined^5,6.     

A Mathematical Technique for Estimating Blastodisc:Yolk Volume Ratios instead of Egg Sizes

Egg size alone is a poor and misleading variable in life-history studies. A mathematical technique for estimating yolk and blastodisc volume ratios in fishes, a much more meaningful character, is generated from first principles. The technique is demonstrated with an example of early ontogeny in fishes of the genus Lucania (Pisces: Cyprinodontidae). Wild, adult rainwater killifish, Lucania parva, and bluefin killifish, L. goodei, were collected in Florida and transported to the laboratory, where offspring were reared under controlled conditions. Offspring were sampled at the onset of cleavage, for simple measurements of yolk and blastodisc morphology. Application of mathematical equations allowed estimates of yolk and blastodisc volumes in the two species. No significant differences were found in clutch size, blastodisc volume, or egg density; however, significant differences existed in the absolute yolk investments, and blastodisc:yolk volume ratios. These differences in reproductive investment within the genus Lucania are interpreted by the altricial-precocial life-history model as a possible causal mechanism in the evolution of species within this genus. The mathematical equations presented in this study enabled us to partition reproductive investment into components that are more biologically meaningful than simple egg size.     

两栖类皮肤石蜡切片样本量探讨——以峨眉髭蟾毛细血管丰富度为例

石蜡切片是量化两栖动物各种皮肤特征的常用技术,但在两栖动物中缺乏取样标准。血管密度等无法直接度量的特性可能需要较大的样本量,探讨这类特征的取样标准有利于实践工作。本研究对雄性峨眉髭蟾Leptobrachium boringii表皮下毛细血管丰富度进行量化,并通过重抽样分析该特征所需样本量。实验获得2只个体10个部位数据,每个部位模拟29种抽样。结果显示,体型较小个体多数部位需接近20条组织或更高的样本量,体型较大个体多数部位所需样本量接近15条或更高。建议在两栖类皮肤石蜡切片工作中对全部有效切片进行制片,以满足不同特征对样本量的要求。表皮下毛细血管丰富度与所需样本量呈负相关关系,可能缘于高密度部位的血管分布更均匀。如在其他两栖动物类群中亦观察到该相关性,则有助于在量化该特征时合理选取样本量。     

Estimating trophic link density from quantitative but incomplete diet data

The trophic link density and the stability of food webs are thought to be related, but the nature of this relation is controversial. This article introduces a method for estimating the link density from diet tables which do not cover the complete food web and do not resolve all diet items to species level. A simple formula for the error of this estimate is derived. Link density is determined as a function of a threshold diet fraction below which diet items are ignored ("diet partitioning function"). Furthermore, analytic relationships between this threshold-dependent link density and the generality distribution of food webs are established. A preliminary application of the method to field data suggests that empirical results relating link density to diversity might need to be revisited.     

手持密度仪检测鸡与鸭精子密度技术的研究

精子密度仪在哺乳动物中已推广应用,但在家禽中还研究很少。本文以黑凤鸡和攸县麻鸭精液为实验材料,手持精子密度仪与血细胞计数板为检测工具,对影响精子密度测定方法准确率因素进行分析,建立鸡、鸭精子密度-吸光度函数并进行验证。结果表明:用密度仪测定时的稀释液(简称"测试液")为3%NaCl时,精液稀释后应尽快检测吸光度;样液在比色皿中的混匀方式对吸光度测定有很大影响;精液用3%NaCl以及3种常用家禽精液稀释液进行10倍稀释后测定吸光度,B液吸光度显著高于与其它3组(P<0.05),其它3组间差异不显著(P>0.05)。用测试液为3.0%NaCl,鸡和鸭精子密度-吸光度呈三次函数关系,回归方程分别是Y_1=1.374X^3-0.786X^2+0.945X-0.002(R^2=0.997)、Y_2=1.283X^3-0.899X^2+0.994X-0.009(R^2=0.996);用0.9%NaCl代替3%NaCl作测试液简化精子密度测定方法可行,鸡精子密度-吸光度回归方程为Y_3=-0.264X^3+1.23X^2+0.468X+0.019(R^2=0.999)。鸡精液测试液为0.9%NaCl、3%NaCl时两种函数的吸光度最佳范围分别是0.035～0.692、0.069～0.624,在此范围内计数板与方程两种方法得到的密度差异不显著(P>0.05),以计数法为真实值,两种方法平均相对误差分别为6.95%、3.11%。以3%NaCl为测试液,鸭精液函数所测样品吸光度最佳范围小于鸡精液的,以计数板法为真实值,在吸光度0.054～0.123范围内的,函数与计数板两种方法所得的精子密度的平均相对误差为4.61%。本文将促进家禽手持精子密度仪研发,以及更好的使用哺乳动物精子密度仪。     

薄层色谱法鉴定天麻胶囊中的天麻素

为了建立天麻胶囊中主要有效成分天麻素的快速鉴定方法,根据天麻素的理化特性,使用乙醇和甲醇提取天麻胶囊中的天麻素,使用薄层色谱法进行鉴定,并与高效液相色谱法的分析结果进行比较。结果表明,薄层色谱法的鉴定结果与高效液相色谱法的检测结果一致,能较准确地鉴别天麻胶囊的真伪。本研究结果表明薄层色谱法能快速简便、准确灵敏地检测天麻胶囊中的有效成分,可作为法定鉴定方法的补充,对天麻胶囊实施快速初筛。