Morphometric study of human alveolar ducts based on serial sections

Interactive computerized morphometry was used for the quantitative study of the terminal airway branches (alveolar ducts) that followed the last bronchioles in three human acini. Two normal adult human lungs from the autopsy service were fixed by instillation and serial sections were prepared; three tissues blocks showing a central bronchiole were selected. Primary and secondary alveolar walls were traced and the following parameters were measured: volume, surface area (of primary and secondary septa), curvature (in equivalent radius) for branches of individual generations, and cumulative values starting with the first alveolar duct and moving peripherally. Although branching was dichotomous, we noticed considerable asymmetry in the pattern of branching and number of side branches. The branching trees of alveolar ducts that we studied comprised 6,7, and 10 generations. The average volume of ducts was 0.04-0.13 mm3, the surface area of primary walls ranged from 0.3616 to 0.7931 mm2 and of secondary septal walls from 0.0100 to 0.0647 mm2. The equivalent radius of curvature was between 22.7 and 38.1 microns. Cumulative increases of volume and surface area revealed similarity in the first five generations. Secondary walls represented only 4% (or 8% if 2 sides are considered) of the primary surface area, strengthening the view that alveoli are incompletely developed side chambers secondary to the alveolar ducts.     

Cytophotometry of DNA in serial sections]

DNA of leprosy cells in serial paraffine sections stained after Felgen were studied photometrically in scanning microscope SPM with electron computer Wang 720C. Significant differences in extinction values (up to 30--35%) between individual sections within the same series were revealed. Cumulative results of different series were rather uniform. It was concluded that photometric studies of serial sections should be carried out for reliable determination of DNA.     

Chronobiologic serial sections of several orders

The chronobiologic serial section is described. Its applicability to the analysis of nonequidistant data is emphasized. Its ability to detect and quantify multiple components is discussed and exemplified on simulated series with various amounts of additive Gaussian noise. This least-squares method is discussed in the context of a number of complementary procedures such as complex demodulation and linear-nonlinear least-squares rhythmometry.     

Interpretation of electron micrographs of single and serial sections

A method of securing serial sections for electron microscopy is described. Serial sections present certain anomalies of interpretation of a nature such that a complete and detailed three-dimensional reconstruction of the sectioned tissue cannot be made. These anomalies are discussed, as well as those which have been encountered in the interpretation of single sections. Observations of the following kinds have been made in an attempt to elucidate the interpretation of single and serial sections: differing methods of mounting adjacent sections, observation of the same section by high-angle stereoscopy, and examination of sections which have been shadowed prior to and subsequent to electron microscopy. It is found that the appearance of sections is independent of the choice of side to be placed against the formvar films. Stereoscopy shows that the appearance of fine structures is strongly dependent upon the direction of the penetrating electron beam with respect to the plane of the structures. Stereoscopy, combined with shadowing, shows quantitatively that extensive sublimation of polymer occurs upon normal exposure in the electron microscope. Observation of sections shadowed prior to electron microscopy indicates that varying amounts of material are removed between sections by the action of microtomy; i.e., it is probable that the sum of the thicknesses of several serial sections is considerably less than the total thickness of material removed from the block. It is believed that this effect, combined with the effect of sublimation, aids in explaining the failure of adjacent sections to exhibit continuity in their detailed structures.     

Three-dimensional computer reconstructions from serial sections of cell nuclei

The analysis of ultrathin serial sections as 3-dimensional (3D) information requires interpretation and display of a large amount of data. We suggest a simple way to solve this problem; it permits presentation of a series of sections as a 3D color image of good quality. It involves a picture system with specialized hardware and software written for this purpose. 3D images of cellular organelles have been drawn either by manually defining the contour of the objects or by thresholding of the volumes in the structures. These 2 methods allow rapid drawing of the image on the screen. It is possible to determine the position, shape and size of 3D structures. This interactive system allows the user to choose between several options: colors, removal of parts of the object, and cutout.     

Computer-aided three-dimensional reconstruction of nematode embryos from EM serial sections

Embryos of the nematode Caenorhabditis elegans were serially sectioned and photographed in the electron microscope (EM). The micrographs were used to produce three-dimensional (3D) reconstructions. Size and position of each nucleus were entered into a computer, displayed as spheres, and were color-coded to indicate lineage membership. Location in space and position in the cell cycle are generally adequate criteria to identify cells. The reconstructions allow visualization of lineage-related topographic patterns and ultrastructural analysis of differently determined cells.