摘要:

AbstractThe development of a quantitative structural platform for experimental biology—extending across a hierarchy of sizes ranging from molecules to organisms—has been punctuated by a series of major achievements over the last 30 years. Stereology, a form of quantitative morphology, has contributed handsomely to this success. A personal view is presented highlighting key events in the development of biological stereology. We also examine stereology with a view toward future developments in biology and speculate how stereology might contribute to the new biological infrastructure currently being built with computers. © 1992 Wiley‐Lis

ISSN:1059-910X    

DOI:10.1002/jemt.1070210402

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThe use of computers in morphometry can involve (1) automated image analysis, semiautomated image analysis and point, intersection, intercept and profile counts of two‐dimensional images on tissue sections with mathematical extrapolation to the third dimension, (2) direct measurement of volumes, surfaces, lengths, and curvature using x,y,z coordinates of serial sectioned images, or (3) stereologic techniques and serial sections which is a combination of 1 and 2 above. Automated and semiautomated image analysis are generally restricted to specimens that are characterized by differential contrast such as interalveolar septa in the lung or histochemically stained mucous granules in pulmonary epithelium. Point, intersection, and profile counts using hand‐held, notebook PCs, portable PCs, or standard PCs and MS‐DOS—based application programs are extremely efficient, precise, affordable, and convenient methods of quantitating average values of a population. When morphometric measurements of individual structures are required, computer‐assisted three‐dimensional reconstruction using x,y,z coordinates of the surface outline from serial sections is a tedious yet precise method. We describe a computer program that efficiently estimates mean caliper diameter, volume, and surface area with less than five percent error with five sections per structure. We also describe a program that does digital image subtraction on serial sections, superimposes digitally generated test systems on biological images, and accumulates point, intersection, and profile counts using a Macintosh II series computer. © 1992 Wil

ISSN:1059-910X    

DOI:10.1002/jemt.1070210403

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractWe describe the application of automatic image analysis for quantitative morphological studies of peroxisomes in rat liver. For automatic detection by light and electron microscopy peroxisomes must be stained with the alkaline DAB procedure for catalase. There is a good agreement between the results obtained by conventional morphometric techniques and by automatic image analysis of DAB‐stained electron microscopic preparations. Moreover, the image analyzer may be used in conjunction with a light microscope for evaluation of semithin sections (1–0.25 μm), provided the section thickness factor is taken into consideration. This latter approach has proven highly efficient in estimation of peroxisome proliferation. The limitations of this method and the relevance of volume density as a reliable morphometric parameter for evaluation of peroxisome proliferation are discussed. In the second part of this study we present the application of image analysis for quantitation of alterations of individual peroxisomal enzyme proteins after treatment with bezafibrate in immunogold stained ultrathin sections. There is good agreement between the results of quantitative immunocytochemistry and Western (immuno) blot analysis of highly purified peroxisomal fractions. In our experience quantitative immunoelectron microscopy provides a versatile, highly sensitive, and efficient method for detection of modulations of various proteins in peroxisomes. Finally the limitations and prospects of quantitative immunocytochemistry for investigation of peroxisomal proteins are discussed. © 1992 Wiley‐Li

ISSN:1059-910X    

DOI:10.1002/jemt.1070210404

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThis study examines the feasibility of combining computer image digitization, image enhancement, and point counting stereological techniques to quantify video images from transmission electron microscopes (TEM). The essential hardware consists of an IBM PC/AT, a Matrox imaging board, a digitizing tablet, a high resolution black and white monitor, and a portable mass storage device. In addition a video camera must be mounted to the TEM. The software is written in three modules which have numerous routines for image acquisition, enhancement, and quantification. Quantification is achieved by selecting an electronic lattice and superimposing it on the cell image. A cursor is moved on the lattice (via the digitizing tablet) and the points are entered into a spreadsheet. One of the major limitations of the system was the reduced resolution inherent in the current hardware. However, sampling experiments showed that one could compensate for the reduced resolution by increasing the magnification of the digitized images, and the stereological values from digitized images compared favorably to those from electron micrographs. Furthermore, the system proved advantageous by eliminating the usual darkroom work, and in enhancing low contrast tissue. In spite of several hardware limitations, the concept of quantifying computer digitized TEM images appears promising. © 1992 Wiley‐Liss, I

ISSN:1059-910X    

DOI:10.1002/jemt.1070210405

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThis paper presents a snapshot view of the influence and direction of microcomputer technology for image analysis techniques in diagnostic pathology.Microcomputers have had considerable impact in bringing image analysis to wider application. Semi‐automated tracing techniques are a simple means of providing objective data and assist in a wide range of diagnostic problems. From the common theme of reducing subjectivity in diagnostic assessment, an extensive body of research has accrued. Some studies have addressed the need for quality control for reliable, routine application.Video digitizer cards bring digital image analysis within the reach of laboratory budgets, providing powerful tools for investigation of a wide range of cellular and tissue features. The use of staining procedures compatible with quantitative evaluation has become equally important. As well as assisting scene segmentation, cytochemical and immunochemical staining techniques relate the data to biological processes.With the present state of the art, practical use of microcomputer based image analysis is impaired by limitations of information extraction and specimen throughput. Recent advances in colour video imaging provide an extra dimension in the analysis of multi‐spectral stains. Improvements will also be felt with predictable increase in speed of microprocessors, and with single chip devices which deliver video rate processing. If the full potential of this hardware is realized, high‐speed, routine analysis becomes feasible. In addition, a microcomputer imaging system can play host to companion functions, such as image archiving and transmission.With this outlook, the use of microcomputers for image analysis in diagnostic pathology is certain to increase. However, it is the software in both design and concept which ultimately governs the performance which can be achieved. Progress may be made by structured software techniques, by application of mathematical principles, or by use of expert systems for data or image interpretation. © 1992 Wiley‐L

ISSN:1059-910X    

DOI:10.1002/jemt.1070210406

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractQuantitative estimations of perimeter and area from digitized video images, and the application of these features in morphometry, are discussed. Estimations from manual tracings via interactive peripherals and from chain codes are addressed. Topics discussed are calibration, determination of vertical and horizontal pixel resolution, effects of tracing jitter, and for chain codes, the spatial quantization scheme representation of the digital contour. Finally, new perimeter estimators for 4‐connected and 8‐connected chain codes for non‐unity pixel aspect ratio are presented with simulation re

ISSN:1059-910X    

DOI:10.1002/jemt.1070210407

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractMorphometric techniques have been developed to quantitatively characterize groups of transmitter‐identified neuronal profiles, such as cell groups, dendrite and nerve terminal fields. These morphometric techniques will be illustrated by introducing some general tools for image analysis which can be considered as a background for the present specific applications. The following methods have been included: (1) methods to identify and quantitatively characterize, from both numerical and geometrical standpoints, groups of profiles in a two‐ and three‐dimen‐sional frame; (2) methods to evaluate the evenness of a certain distribution of profiles in the plane; (3) methods to identify subgroups of profiles based on their different spatial or optical density; and (4) methods to compare the distributions of two or more groups of profiles. The applications of these general tools to some neuroanatomical problems, such as cell group definition and description, have been illustrated. Practical examples performed on immunocytochemical preparations of neuronal profile populations are also given. Finally, the potentiality of numerical classification to classify and compare morphometric data has been shown. As an example, numerical classification methods have been applied to the morphometric and microdensitometric analysis of adrenaline/neuropeptide Y costoring neuronal systems of the brainstem in adult and aged rats. © 1992 Wiley

ISSN:1059-910X    

DOI:10.1002/jemt.1070210408

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThis paper describes a computer‐aided tutorial for biological stereology. Stereology, a type of quantitative morphology, includes a collection of statistical methods that quantify the structural compartments that can be viewed in sections with light and electron microscopy. These methods provide volume, surface, length, shape, and number data, and help define the quantitative relationships among the structural compartments of biological hierarchies. Hierarchies, which connect structural data ranging in size from molecules to organs, serve as a central core to which the data of biological databases can be linked. The tutorial focuses on two objectives. It provides the user primarily interested in using quantitative morphology databases with background information, and offers a set of state‐of‐the‐art tools to researchers wishing to use these methods in the laboratory. The main topics of the tutorial include: introduction to quantitative morphology, symbols/terms, data types, sampling, hierarchies, data interpretation, and utilities. The tutorial runs under the MS‐DOS operating system and requires at least an IBM PC AT (or compatible), a color monitor (EGA, VGA), 540 KB of RAM, and 3 MB of hard disk space. © 1992 Wiley

ISSN:1059-910X    

DOI:10.1002/jemt.1070210409

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThe paper describes microcomputer software for point counting stereology. Stereology includes a collection of statistical methods that quantify the images of light and transmission electron microscopy. The methods use test grids placed over images to collect raw data, which includes counts of points, intersections, transections, and profiles. In turn, the counts are included in stereological equations that give estimates of compartmental volumes, surfaces, lengths, or numbers. These parameters describe the composition of a structure in three‐dimensional space. The PCS (point counting stereology) System Software III serves as a data collection, storage, and management tool. Users set up point counting protocols without programming, enter data by pressing predefined function (MS‐DOS) or alphabetic keys (UNIX), store data in files, select files for analysis, and calculate results as stereological densities. The latest version of the PCS software includes a new user interface and is designed as a research “front end” that can feed data either into the calculation tools of a stereology tutorial (Bolender, 1992, this issue) or into the analysis routines of quantitative morphology databases (Bolender and Bluhm, 1992). © 1992 Wiley

ISSN:1059-910X    

DOI:10.1002/jemt.1070210410

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractA new application of techniques for preparing impervious biological specimens for light microscopy (LM) and transmission electron microscopy (TEM) has been developed. Microwave irradiation was used to facilitate fixation. A priming technique was used to increase the bonding of the outer surface of the specimens with the resin. Priming the waxy or cuticular surface with Z‐6040 (gamma‐glycidoxypropyl trimethoxysilane) solved the problem of specimen “pull out” from the resin. Insect specimens with various types of cuticles (waxy or chitinous) and seeds were successfully studied ultrastructurally using this technique. © 1992 Wiley

ISSN:1059-910X    

DOI:10.1002/jemt.1070210411

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY