In this review we present immunohistochemical methods for visualization of capillaries and muscle fibres in thick muscle sections. Special attention is paid to the procedures that preserve good morphology. Applying confocal microscopy and virtual 3D stereological grids, or tracing of capillaries in virtual reality, length of capillaries within a muscle volume or length of capillaries adjacent to a muscle fibre per fibre length, fibre surface area or fibre volume can be evaluated by an unbiased approach. Moreover, 3D models of capillaries and muscle fibres can be produced. Comparison of the developed methods with counting capillary profiles from 2D sections is discussed and the reader is warned that counting capillary profiles from 2D sections can underestimate the capillary length by as much as 75 percent. Application of the described 3D methodology is illustrated by the anatomical remodelling of capillarity during acute denervation and early reinnervation in the ra t soleus and extensor digitorum longus muscles., I. Eržen, J. Janáček, L. Kubínová., and Obsahuje bibliografii a bibliografické odkazy
A simple, vector-based graphics editor, designed for experimental data presentation, schematics, large structured chemical formulae etc. in a modest IBM PC, MS DOS environment is described. Its applications in semiautomatic image analysis, especially ¡n the implementation of design-based stereological methods for estimating global geometrical characteristics of studied samples and in 3D volume reconstruction from serial sections are discussed.
The STESYS2 software is a new version of the IBM PC software supporting interactive stereological measurements. In comparison with the previous STESYS, it is enhanced by a number of useful options, e.g. on-line image input via a TV camera coupled with a microscope operating under MS Windows OS. The main advantage, when compared with other such software packages, is the design of the STESYS2 as a module of the freeware image processing system Image Tool which provides a user-friendly environment including a number of image processing and preprocessing routines. Capabilities of the STESYS2 are illustrated by a practical example: estimation of the surface area of capillaries in the terminal villi of human placenta by the Sandau spatial grid method., Z. Tomori, L. Matis, P. Karen, L. Kubínová, I. Krekule., and Obsahuje bibliografii
Measurements of ultrastructural characteristics of chloroplast thylakoids are important for studies of ontogenic or ecological limitations of leaf photosynthetic functions. Most frequently, volumetric proportion of thylakoids in the chloroplast is measured; however, such measurement does not provide a direct information about the surface area of thylakoids which is most important from the functional point of view. Therefore, we adapted the stereological method using "local vertical windows" for estimating thylakoid surface area in the chloroplast volume and compared thus obtained surface density results with results of conventional volume density measurements. The methods were tested in the study of chloroplast ultrastructure in the leaves of plants of two maize (Zea mays L.) hybrid combinations, 2013×CE810 and CE704×CE810, developing in control and chilling conditions. Correlation analysis revealed a tight relationship between the granal/intergranal thylakoid surface density and volume density results, both indicating that under chilling conditions the development of the system of thylakoids in maize leaves is suppressed, while the difference is more pronounced in CE704 than in CE810 genotype, known to have a better photosynthetic performance. and L. Kubínová, J. Kutík.
Changes in Hill reaction activity (HRA) and ultrastructure of mesophyll cell (MC) chloroplasts were studied during the ontogeny of third leaf of maize plants using polarographic oxygen evolution measurement, transmission electron microscopy, and stereology. The chloroplast ultrastructure was compared in young (actively growing), mature, and senescing leaves of two different inbreds and their reciprocal F1 hybrids. Statistically significant differences in both HRA and MC chloroplast ultrastructure were observed between different stages of leaf ontogeny. Growth of plastoglobuli was the most striking characteristic of chloroplast maturation and senescence. The chloroplasts in mature and senescing leaves had a more developed system of thylakoids compared to the young leaves. Higher HRA was usually connected with higher thylakoid volume density of MC chloroplasts. and J. Kutík ... [et al.].
Influence of moderate chilling stress on vascular bundle sheath cell (BSC) and especially mesophyll cell (MC) chloroplasts of mature maize leaves was studied by electron microscopy and stereology. Plants of two inbred lines of maize, differing in their photosynthetic activity, and their F1 hybrids were cultivated during autumn in heated or unheated glasshouse. Generally, chilling temperatures resulted mainly in the decrease in stereological volume density (VD) of both granal and intergranal thylakoids of MC chloroplasts, while the ratio of granal to all thylakoids (granality) was less affected. The VD of peripheral reticulum and plastoglobuli usually increased after cold treatment of plants. The volume of MC chloroplasts usually increased under chilling stress, the shape of the chloroplasts changed only slightly. The ultra-structure of chloroplasts differed between individual genotypes; chilling-stressed hybrid plants showed positive heterosis particularly in the granal thylakoids' VD of MC chloroplasts. and J. Kutík ... [et al.].