Scanning electron microscopy最新文献

Maintenance of tear film in normal conditions is dependent on mucus layer integrity and the presence and distribution of conjunctival epithelial cell microvilli. In the present work a new methodology has been developed to gain correlative information about microprojection assessment and mucus composition, from the same specimen, by Light Microscopy (LM), Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). We have characterized the glycosidic residues secreted by goblet cells in normal human conjunctiva, by means of four lectins (WGA, ConA, PNA and SBA), conjugated with FITC for LM and with colloidal gold for TEM and SEM. The cytochemical reactions were performed on histological sections of paraffin-embedded material and on semithin and ultrathin sections of both Epon embedded material directly processed for TEM and of blocks recovered from SEM and reprocessed for TEM. WGA, ConA, PNA and SBA receptors were found to be constituents of the mucus produced by goblet cells in human conjunctiva. The granules of the so-called "second mucus system" (SMS) cells were labelled mainly by WGA. A difference in the quality of glycoconjugates between goblet cells and SMS cells has been also demonstrated. Our results provide an improved method to evaluate alterations of tear film that occur in many conjunctival diseases.

The organization of cytoskeletal proteins in whole-mount adherent platelets from African green monkeys and normal human volunteers has been studied by SEM, high vacuum electron microscopy (HVEM) and conventional (120 kV) electron microscopy. We describe three distinct organizational zones, the Central Matrix, the Trabecular Zone and the Peripheral Web in spread platelets from both sources. The Central Matrix is an ill-defined superstructure of 80-100 A filaments of short length which enshrouded the granules, dense bodies, mitochondria and elements of the open-channel and dense-tubular systems. The latter, identified through the use of peroxidase cytochemistry with the whole mounts, is an anastomosing network of elongate saccules having diameters of 600-1200 A. The Trabecular Zone, which encircles the Central Matrix, contains 165, 80-100 and 30-50 A filaments in an open lattice of irregular lattice spacing. The outermost region of the cells, the Peripheral Web, is comprised of 70 A filaments organized in a honeycomb lattice with center to center spacing in the range 150-300 A. This pattern for the spread cells is not consistently observed in cells during the early stages of adhesion; therefore, correlations of SEM and TEM observations are made for the various stages of adhesion/activation.

The elemental content of anagen hair fibers in a Caucasian population of healthy females and males in the age range 10-69 years was performed to constitute a baseline for further studies of pathological conditions. Proton induced X-ray emission (PIXE) analyses were performed on single hair fibers in triplicate from 103 individuals in order to determine sulfur, zinc, calcium, and chlorine content. The hair fibers were all anagen hairs collected from a site little influenced by genetic and hormonal influences 1.5 cm above the right ear of the probands. An area 5-8 mm from the follicle bottom was chosen for the analysis in all cases to minimize effect of hair-do contamination. The average sulfur content was 0.049 g/g and the average zinc content 170 micrograms/g. These results were not significantly influenced by chloroform/ethanol rinsing before analysis. The calcium and chlorine contents were 330 micrograms/g and 0.0033 g/g respectively. The latter data are expected to be more seriously influenced by external factors (e.g., contamination) than sulfur and zinc. No correlation between elemental concentration and sex was found for sulfur and zinc in the present material. PIXE analysis of single hair fibers yields valuable information on the elemental composition of hair fibers and can be rapidly and efficiently performed after simple mounting procedures.

The testes of some different orders of eutherian mammals were examined by conventional electron microscopy with respect to their pattern of spermiogenesis. In addition, some of the testes were studied by cytochemical methods for demonstration of certain nuclear proteins and of glycoproteins in the acrosome and the plasma membrane of spermatids. It was found that although the basic pattern of spermiogenesis was similar in all species studied, there were pronounced dissimilarities in the final shape of the spermatids. Differences were also observed in the timing of the differentiation of several organelles. The head of late spermatids and spermatozoa of Primates, Carnivora and Perissodactyla was cone-shaped, whereas in Artiodactyla and Lagomorpha it was flattened or paddle-shaped, and in Rodentia hook-shaped. The size and shape of the acrosome varied considerably between the orders, as did the length of the middle piece.

A patient who repeatedly produced urinary calculi, had consumed about 3 g of cristobalite (SiO2) per day for many years. Investigations using scanning electron microscopy revealed minute particles containing silicon in the core of the stone as well as in urine sediment. A mechanism similar to that proposed for the effect of silicon-containing drugs against gastric ulcer, may play a role in this formation of silicon-containing urinary stones.

This paper reviews ultrastructural studies of the intercellular contacts or junctions between cells of the bone marrow. Studies using tannic acid and glutaraldehyde as a fixative have shown pentalaminar complexes between many types of cells in marrow of mice and chicks. These intercellular contacts occur between adjacent stromal cells, between stromal cells and developing blood cells and, in marrow of mice, between migrating blood cells and cells of the sinusoidal wall. Because of their location and widespread occurrence, it is believed these contacts may represent a type of adherent junction helping to maintain an orderly arrangement of blood cells and stromal cells in the marrow. Migrating blood cells may use these contacts as anchoring sites during locomotion toward the sinusoids and in crossing the sinusoidal wall. On the other hand, since these junctions resemble gap junctions of other tissues, one should not exclude the possibility that they are involved in cellular communication. Freeze-fracture and lanthanum impregnation studies have failed to demonstrate these junctions in marrow. Studies using ruthenium red have shown apparent sites of attachment between cells of the marrow, but it is not known whether these sites correspond to the intercellular contacts seen in tannic acid preparations.

We have made several observations during the course of our studies that show the presence of matrix effects in soft biological tissue and standards. The sputtering rate of gelatin is approximately twice that of epoxy resin, but the ion yield of lithium in gelatin is an order of magnitude less than in epoxy. Osmium impregnation of freeze-dried material significantly alters the localization of calcium, but not potassium and barium. The absolute count rate for calcium in osmicated tissue is increased several-fold above that in freeze-dried tissue. Scanning electron microscopy of sputtered material shows the formation of cones during sputtering, which is particularly, but not exclusively, associated with melanin granules and red blood cells. These structures are known to be highly emissive for Na, K, and Ca. Boron implanted tissue also exhibits selective boron emission from melanin granules. Relative proportions of monoatomic and polyatomic emission vary in epoxy, gelatin and tissue. Ion images of carbon, chlorine and vanadium in tissue embedded with a vanadium-doped epoxy resin show variations in local regions that correspond to tissue structure. The energy distributions of common secondary ions differed somewhat in resin and two different tissue regions. These examples show the existence of potential matrix effects in soft biological tissue that involve both differential sputtering and ion yield effects.

Thebesian vasculature provides for communication between the coronary system and the chambers of the heart. Anatomic, embryologic, physiologic, and therapeutic investigations have involved this component of cardiac anatomy from the early 18th century to the present time. The scanning electron microscope (SEM) now affords an innovative approach to the study of the ostia of these veins as they open into the chambers of the heart. The surface of the intact endocardium is continuous, whether it is treated with boric acid or not, as long as it remains intact. Enzymatic microdissection of tissues with trypsin, hyaluronidase and pronase, followed by similar treatment with boric acid, reveals continuity of successive component layers of the endocardium extending into Thebesian substructure. Thebesian tributaries are easily visualized from the ostia but the deeper capillary network of the Thebesian system is not demonstrable by this approach. Valvular structures such as might prevent retroflow during the cardiac cycle are not present. Our observations with SEM support anatomic relationships indicated by previously published work.

The renal microvasculature (afferent arteriole) and glomeruli were examined and quantitated by two methods in the post-ischemic hydronephrotic (PIH) kidney. The methods used were: an in vivo examination and controlled perfusion-fixation, quantitative vascular casting examined by scanning electron microscopy. The second method was also applied to the vasculature of the contralateral, functional kidney. The goals of the study were to: validate the quantitative vascular casting method by comparing PIH renal microvascular data from the casting method with in vivo values and determine the extent of microvascular dimensional difference of the PIH kidney from its contralateral functional counterpart. It was determined that the casting values were consistent with the data obtained from the in vivo examination of the afferent arteriole and glomeruli. This finding provides further support for the quantitative renal microvascular casting technique. Using that technique it was determined that the dimensions of the microvasculature and glomeruli of the PIH kidney were severely (and significantly, p less than 0.05) reduced compared to its functional mate. Since these PIH vessels show a significant decrement in size, vascular reactivity and functional data based on the PIH vessels should be looked at cautiously. The vasculature and glomeruli of the PIH kidney might not be totally normal, however structurally, the glomeruli do not appear to be dramatically altered.

Vascular corrosion casts of Lewis lung carcinomas (LLC) grown subcutaneously in C57BL/6-mice are correlated with histological sections and with tumor tissue prepared for scanning electron microscopy (SEM). By making low, medium and high pressure cast preparations we studied the influence of perfusion and injection pressure on the resulting cast sample. Three types of vascular proliferations are distinguishable in LLC: Small globular outgrowths on sinusoidal dilated tumor capillaries, caused by proliferation of their endothelial cells. New sprouts on surrounding host vessels, invading the small, still avascular implant. Superficially located, centrifugally running sprouts in peripheral regions of large tumors. They invade the surrounding host tissue. Vascular sprouts are of venous origin, have a fragmentary endothelium and are rather "leaky" if casted. High pressure preparations of large tumors reveal central avascular cavities surrounded by centripetally running, compressed and blind ending tumor vessels. Irrespective of the applied injection pressure, the casts always exhibit extravasal channels caused by degeneration of the endothelium of central tumor vessels. We show that SEM of vascular corrosion casts combined with histology not only demonstrates such contrary processes as the development of tumor blood vessels and the simultaneously occurring vascular degeneration, but also elucidates all other morphological characteristics of the tumor vascular system.