Erythroid cells of the bone marrow of the mouse

The bone marrow of three intact male mice of C57Bl/6 line, fixed by perfusion of isotonic fixative of Karnovsky, has been studied by means of the scanning electron microscopy method. The surface of erythroid cells, that are immediately connected with macrophages of the erythroblasts islets, is analysed. According to the surface form, the erythroid cells are devided into 5 types. Every maturation stage of the erythroid cells is characterized by a certain type of surface. For identification of basophilic and polychromatophilic proerythrocytes the combined method of light and electron scanning microscopy of the cells in suspension is used. The bone marrow cells, obtained from the two male mice of C57Bl/6 line are fixed with the same fixative on special glasses with grids traced on them, stained after Romanovsky-Giemsa method and in moist preparations are examined in the light microscope. After further treatment the surface of the same cells in studied in the scanning electron microscope.     

The use of backscattered electrons in analytical electron microscopy for the measurement of the mass of individual rat blood platelets

Summary The backscattered electron signal, generated in individual cells, has been used to measure the dry mass of these cells. Absolute mass values were obtained by comparing the backscattered electron signals of cells to the signals of polystyrene-latex spheres of known mass. The technique was carried out in an automated analytical scanning transmission electron microscope and applied to rat blood platelets.The resulting mass distributions agreed well with the distribution measured with a method that uses the transmitted electron signal by means of densitometric analysis of electrographs. Also the range of masses was in agreement with values deduced from data in the literature.The fully automated technique has the advantage that it is direct, fast, and that thicker specimens can be measured than is possible using the transmitted electron signal. The method is intended for use in combination with quantitative electron probe X-ray microanalysis and is then able to produce elemental mass fractions of biological specimens at the subcellular level.In honour of Prof. van Duijn     

The use of backscattered electrons in analytical electron microscopy for the measurement of the mass of individual rat blood platelets

The backscattered electron signal, generated in individual cells, has been used to measure the dry mass of these cells. Absolute mass values were obtained by comparing the backscattered electron signals of cells to the signals of polystyrene-latex spheres of known mass. The technique was carried out in an automated analytical scanning transmission electron microscope and applied to rat blood platelets. The resulting mass distributions agreed well with the distribution measured with a method that uses the transmitted electron signal by means of densitometric analysis of electrographs. Also the range of masses was in agreement with values deduced from data in the literature. The fully automated technique has the advantage that it is direct, fast, and that thicker specimens can be measured than is possible using the transmitted electron signal. The method is intended for use in combination with quantitative electron probe X-ray microanalysis and is then able to produce elemental mass fractions of biological specimens at the subcellular level.     

Decrease of NADH in HeLa cells in the presence of transferrin or ferricyanide

The short-term incubation of HeLa cells in the presence of diferric transferrin or ferricyanide, which are reduced externally by the transplasma membrane reductase, produces a stoichiometric decrease in NADH and increase in NAD+, which is stimulated by insulin. The NADP/NADPH ratio does not change during 15 min incubation with the oxidants. The total pyridine nucleotide pool of HeLa cells is not affected. Incubation with apotransferrin and ferrocyanide, which cannot act as oxidants for transmembrane electron transport, does not change the pyridine nucleotide concentrations in the cells. Our results show that NADH can act as the internal electron donor for the reduction of external oxidants by the transmembrane reductase. It appears that oxidation of NADH by the transmembrane electron transport using ferricyanide or iron transferrin as external electron acceptors is sufficient to stimulate growth in HeLa cells.     

Ultrastructure of the surface of the iris in the rat eye

Summary The surface structure of the iris in the rat eye was studied by light and electron microscopy.The anterior surface of the rat iris is covered with a discontinuous layer of large, polygonal endothelial cells with microvilli on their surface. Crypts and holes between adjacent endothelial cells extend into the stroma and form there a complicated network of interconnected spaces occupying about one half or more of the volume of the pupillary part of the stroma. The crypts are occasionally partly covered with endothelial cells. The posterior surface is covered with a continuous layer of polyhedronal epithelial cells. These are covered with many folds and processes, partly masked by an amorphous coat. The sphincter pupillae and dilatator muscles are possible to recognize on the scanning electron micrographs as well as blood vessels and nerve fibers in the iris stroma.The endothelial cells show many structural similarities with the endothelial cells on the cornea, probably reflecting their common origin. The results obtained, especially those from the scanning electron microscopic studies, are discussed and interpreted in relation to previous studies. The advantages in using different light and electron microscopic techniques are stressed.Supported by grants from Magnus Bergwall's Stiftelse and the Swedish Medical Research Council (B71-12X-2543-03).     

Ultrastructure of the gut neurotensin cell

Summary In mammals, neurotensin cells occur scattered in the epithelium of the jejunum-ileum. In chicken, neurotensin cells are abundant in the region of the gizzard-duodenal junction (antrum) where they occur intermingled with numerous somatostatin and gastrin cells. The neurotensin cells in chicken, dog and man were identified at the electron microscopic level by immunocytochemistry, using the consecutive semithin/ultrathin section technique. They contain numerous electron dense cytoplasmic granules, predominantly in the basal portion of the cell. It was shown that these granules are the storage site for neurotensin. The neurotensin granules are round, highly electron dense and of about the same size in the different species examined (mean diameter 260–290 nm). in dog and man the granules have a tightly applied surrounding membrane while in the chicken a relatively electron lucent zone separates the electron dense core from the granule membrane. The ultrastructure of the neurotensin granules in chicken is some-what reminiscent of that of the gastrin granules. The mean diameter of the gastrin granules in chicken antrum is 230 nm; for the somatostatin granules the mean diameter is 305 nm.     

The histochemistry and ultrastructure of the clitellum of the enchytraeid Lumbricittus rivalis (Oligochaeta: Annelida)

The monostratified clitellar epidermis of Lumbricillus rivalis consists of supporting cells, granular secretory cells, and globular secretory cells, together with the acid mucous cells typical of normal skin in the anterior and posterior transition zones. The secretion of the granular cells is a neutral glycoprotein with low levels of bound lipid, and that of the globular cells is a sulphated acid mucopolysaccharide lacking detectable protein or lipid.
Ultrastructurally the granular cells possess 1 μm, membrane-bound granules of variable electron density suggestive of maturation changes. The formative granules arise from Golgi vesicles and are moderately electron translucent. They contain parallel-aligned, tubule-like inclusions of 14 nm diameter, observed also in the mature regions of the Golgi. Granules in the mid or apical part of the cell show increased electron density, the formative pattern being wholly or partially obscured and the subunit alignment, when discernable, no longer uniform. Moiré fringe patterns are evident in some granules. The patterns and electron density are lost after pronase digestion. The globular cells contain electron translucent, membrane-bound globules of 2–3 μm diameter with finely fibrous contents. They arise from mature Golgi vesicles and are unaffected by pronase treatment.
The results suggest that the granular cells secrete the cocoon wall and the globular cells the material that surrounds the developing embryos, and are compared with published accounts of other microdrile clitella and with those of the clitellum of lumbricid earthworms.     

Ultrastructure of the APUD-like cells of the frog thymus gland

Within the thymus gland of the European common frog, Rana temporaria, cells with endocrine- like appearance have been found. At the ultrastructural level the most characteristic feature of their cytoplasm is the presence of secretory granules. Some cells possess irregular electron lucent granules with an eccentrically located dense core while others possess smaller electron dense granules. The cytoplasm contains also cisterns of rough endoplasmic reticulum, free ribosomes, and small mitochondria. The cells possess irregular nuclei with the pronounced nucleoli. These endocrine-like cells are connected by desmosomes with neighbouring non-granulated epithelial cells. Ultrastructural features of the cells described here resemble those seen in polypeptide hormone-secreting cells belonging to the family of cells of the APUD (Amine Precursor Uptake and Decarboxylation) series.     

Ultrastructure of the gut neurotensin cell.

In mammals, neurotensin cells occur scattered in the epithelium of the jejunum-ileum. In chicken, neurotensin cells are abundant in the region of the gizzard-duodenal junction (antrum) where they occur intermingled with numerous somatostatin and gastrin cells. The neurotensin cells in chicken, dog and man were identified at the electron microscopic level by immunocytochemistry, using the consecutive semithin/ultrathin section technique. They contain numerous electron dense cytoplasmic granules, pre-dominantly in the basal portion of the cell. It was shown that these granules are the storage site for neurotensin. The neurotensin granules are round, highly electron dense and of about the same size in the different species examined (mean diameter 260--290 nm). In dog and man the granules have a tightly applied surrounding membrane while in the chicken a relatively electron lucent zone separates the electron dense core from the granule membrane. The ultrastructure of the neurotensin granules in chicken is somewhat reminiscent of that of the gastrin granules. The mean diameter of the gastrin granules in chicken antrum is 230 nm; for the somatostatin granules the mean diameter is 305 nm.     

Effect of chloramphenicol on the electron systems in Ustilago cynodontis.

The mycelial cells of Ustilago cynodontis possess at least two electron transport systems: a cyanide-sensitive cytochrome pathway, which represents the major route for electron transport, and an alternative cyanide-insensitive pathway, inhibited by salicylhydroxamic acid. In the presence of chloramphenicol in the culture medium, mycelial cells respire only by the alternatuve chain. The stable induced yeast-like cells, obtained by prolonged chloramphenicol treatment of the mycelial cells, respire as the untreated mycelial cells; this result indicates that the phenotypic change induced by chloramphenicol is not related to a modification of the respiratory system.     

Fine structure of the dorsal lingual epithelium of the frog, Rana rugosa

Scanning and transmission electron microscopy was employed to investigate the ultrastructure of the lingual dorsal epithelial cells of the frog, Rana rugosa. The specimens for scanning electron microscopy were prepared by a method that involved osmium postfixation and treatment with acid to remove extracellular material that adhered to the surface of the tongue. Over almost the entire dorsal surface, filiform papillae, consisting of a large number of non-ciliated cells with microridges and a very small number of ciliated cells, were compactly distributed. Fungiform papillae were scattered among these filiform papillae. A round sensory disk was located on the top of each fungiform papilla. Each sensory disk was encircled by a band of ciliated cells. Transmission electron microscopy revealed that a large part of the filiform papillar epithelium was composed of cells that contained numerous electron-dense granules. These cells were coincident with the non-ciliated cells observed by scanning electron microscopy. In these cells, the nucleus was located on the basal side, and the ergastoplasm was well-developed on the basal side of the nucleus.     

Involvement of transferrin in the reduction of iron by the transplasma membrane electron transport system

Nonpermeable electron acceptors can be reduced by a transplasma membrane electron transport system in suspensions of intact cells. Here we report that diferric transferrin is reduced by HeLa S3 cells. The reduction is recorded spectrophotometrically as the formation of the ferrous complex of bathophenanthroline disulfonate. Ferric ammonium citrate can also be used as an electron acceptor, and the presence of low concentrations of diferric transferrin greatly stimulates the reduction of trivalent iron under these conditions. Likewise very low concentrations of ferricyanide, which does not give rise to a ferrous bathophenanthroline disulfonate complex formation, have a strong stimulatory effect on the complex formation when ferric ammonium citrate is the source of ferric iron. Apotransferrin is a potent inhibitor of the reaction. The inhibition occurs at the concentration necessary for complete occupancy of the transferrin receptors. The inhibition can be demonstrated also when high concentrations of ferricyanide are used as electron acceptor. The possible mechanism behind the reported phenomena is discussed, and it is concluded that the transplasma membrane electron transport system can be involved in the process of cellular iron uptake.     

Use of electron microscopy to characterize the surfaces of flocculent and nonflocculent yeast cells

The surfaces of flocculent and nonflocculent yeast cells have been examined by electron microscopy. Nonextractive preparative procedures for scanning electron microscopy allow comparison in which sharp or softened images of surface details (scars, etc.) are the criteria for relative abundance of flocculum material. Asexually flocculent budding-yeast cells cannot be distinguished from nonflocculent budding-yeast cells in scanning electron micrographs because the scar details of both are well resolved, being hard and sharp. On the other hand, flocculent fission-yeast cells are readily distinguished from nonflocculent cells because fission scars are mostly soft or obscured on flocculent cells, but sharp on nonflocculent cells. Sexually and asexually flocculent fission-yeast cells cannot be distinguished from one another as both are heavily clad in "mucilaginous" or "hairy" coverings. Examination of lightly extracted and heavily extracted flocculent fission-yeast cells by transmission electron microscopy provides micrographs consistent with the scanning electron micrographs.     

Fine structure of the parotid and mandibular glands of the cotton rat (Sigmodon hispidus).

The parotid and mandibular glands of the cotton rat were examined by light and transmission electron microscopy. Parotid gland: Acinar cells were serous in nature, and contained electron-dense granules. Intercalated duct cells contained electron-dense granules. Striated duct cells had small granules of moderate and high electron densities. Mandibular gland: Acinar cells were seromucous in nature, and contained granules of low and moderate electron densities. Intercalated duct cells contained granules of moderate and high electron densities. Striated ducts were comprised of two portions - a secretory portion and a striated portion without granules. The secretory portion had many electron-dense granules. A sexual dimorphism was obserbed in these granules, which were smaller and fewer in females than in males.     

Some observations on the formation of the primary tunica albuginea in the ovary of the chick embryo (author's transl)

The ovarian cortex of chicken embryos of 10 to 14 days of incubation was observed in order to study the formation of the primary tunica albuginea and to discern in its cells an ultrastructural confirmation of an eventual steroidogenic capacity which has been proposed by some authors. From observations at the electron microscope, it is probable that the mesenchyme-like cells which constitute the primary tunica albuginea may originate from the somatic cells of the cortex, either from those of the germinal epithelium, or from those which, because of their position, may be termed prefollicular. There is not apparent ultrastructural confirmation of an active steroidogenic capacity in the albugineal cells. A possible physiological difference between clear cells and dark cells due to a different electron density, is furthermore discussed.     

Ultrastructural characteristics of the salivary glands of the taiga tick, Ixodes persulcatus (Ixodidae). I. The granulosecreting alveoles of the fasting female

Two types of granulosecreting alveoles were found in salivary glands of hungry females by means of electron microscopy of ultrafine sections. Alveoles of the IInd type occur in the anterior helf of the gland. They are not numerous and consist of three types of secretory cells (A, B, C) surrounding the inneralveolar cavity. The secretory cells are separated from each other and from the basal membrane by the strands of the epithelial cells P. Three types of spherical inclusions were found in the secretory cells. They differ in size, electron density and intensity of staining of half-fine sections with toluidin blue. The apical cytoplasmatic membrane of secretory cells bears numerous microvilli. Alveoles of the IIIrd type, which constitute the main mass of the gland tissue, have a narrow slit-like inneralveolar cavity. The basal part of the alveole is formed by 3--4 large cells filled with large spherical electron-transparent vacuoles of the secretion. The apical part of the alveole is occupied by 9 to 11 cells E, whose cytoplasm is filled with numerous flat cisternae of granular endoplasmatic reticulum and small and medium secretory vacuoles of different electron density. Alveoles of the IInd and IIIrd type of I. persulcatus are not identical with those of Hyalomma asiaticum, Boophilus microplus and other members of the subfamily Amblyomminae.     

Structure of the cell surface of the yeast Candida tropicalis and its relation to hydrocarbon transport

The surface structure of the hypdrocarbon-utilizing yeast Candida tropicalis was investigated by scanning and transmission electron microscopy (SEM and TEM respectively). The sample preparation technique was based on a rapid cryofixation without any addition of cryoprotectants. In subsequently freeze-dried samples the surface structure was analysed by scanning electron microscopy. Thin sections were prepared from freeze substituted samples. Both techniques revealed hair-like structures at the surface of hydrocarbon-grown cells. The hairy surface structure of the cells was less expressed in glucose-grown cells and it was absent completely after proteolytic digestion of the cells. When cells were incubated with hexadecane prior to cyryofixation a contrast-rich region occured in the hair fringe of thin sections as revealed by TEM. Since these structures were characteristic for hexadecane-grown cells and could not be detected in glucose-grown or proteasetreated cells it was concluded that they originate from hexadecane adhering to the cell surface and are functionally related to hexadecane transport. The structure of the surface and its relation to hydrocarbon transport are discussed in view of earlier results on the chemical composition of the surface layer of the cell wall.Abbreviations SEM Scanning electron microscopy - TEM transmission electron microscopy     

The ultrastructural basis for the identification of cell types in the pancreatic islets

Summary The pancreatic islets of rabbit, dog and opossum have been studied by light and electron microscopy. Silver-positive cells in the rabbit are predominantly sandwiched between the peripheral A and central B cells, and by electron microscopy are identified as D cells. Pancreatic islets in the tail of the dog pancreas have A, B, and D (silver-positive) cells, but the islets in the uncinate process of the dog pancreas lack phosphotungstic acid hematoxylin-positive A cells. By electron microscopy the characteristic D cells are found in both tail and uncinate process, but A cells are confined to the tail islets, confirming the identification of cell types. A unique cell type termed the F cell is found in the dog uncinate islets and it is characterized by secretory granules of angular profiles. In the opossum, the A cells contain considerable amounts of glycogen demonstrable by both light and electron microscopy. A unique cell type is also present in the opossum islets termed an E cell (Thomas, 1937), which has large secretory granules (400–500 m). The physiological implications of a multiplicity of cell types in pancreatic islets is discussed.This investigation was supported in part by United States Public Health Service research grants GM-10102 and GM-03784 from the Institute of General Medical Sciences, and AM-01226 from the Institute of Arthritis and Metabolic Diseases. The authors wish to acknowledge the valuable technical assistance of Mrs. Aileen Sevier and Mrs. Lidia Donahue.     

Atypical Granules in the Eyes of the White Mutant of Drosophila melanogaster Are Lysosome-Related Organelles

In the pigment cells of the white mutant of Drosophila melanogaster, as described earlier, two types of abnormal granules are found by conventional electron microscopy. However, both types of abnormal granules, in addition to those in pigment cell invaginations, are also present in the cytoplasm of the photoreceptor cells. Three enzymes (acid phosphatase, peroxidase, and tyrosinase) are localized within the eyes of wild type and white mutant Drosophila melanogaster by electron microscopy. Peroxidase activity is present in lamellar bodies close to the rhabdomeral microvilli of both fly types. However the organelles containing peroxidase activity are 6-fold more frequent in the wild type than in the mutant. Acid phosphatase is present in lamellar bodies between and at the bases of the rhabdomeral microvilli of the wild type, as well as in ommochrome granules of the photoreceptor cells. In the white mutant, however, acid phosphatase was located in electron lucent vacuoles in the cytoplasm of the receptor cells. These acid phosphatase-positive vacuoles also contained both types of abnormal granules. The latter result indicates that abnormal granules in the receptor cells originate from lysosomal degradation and that targeting of lysosomal enzymes is altered in the white mutant. Due to the tyrosinase activity in the hemolymph of flies, the extracellular spaces are electron dense after DOPA incubation. Since some abnormal granules within the photoreceptor cells are not surrounded by an extracellular space, they can be assumed to originate within the photoreceptor cells.     

The fine structure of blood cells in the ascidian Perophora viridis

The fine structure of each of the blood cell types of Perophora viridis has been characterized and strong evidence for localization of vanadium in two of these types is given. There are eight cell types; phagocytes which may contain completely engulfed cells, lymphocytes with a prominant nucleolus and scanty cytoplasm packed with clustered ribosomes, and six other cell types each with distinctive granules. Morula cells contain a central nucleus and cytoplasm filled by wedged bodies, about five of which are seen in section. These bodies contain regularly spaced electron dense foci. Green cells have the same organization but contain bodies which are electron dense throughout. Granular amoebocytes contain many smaller lightly staining oval bodies and much glycogen. Another cell type (probably orange cells of light microscopy) contains numerous granular rounded bodies. Compartment cells have vacuoles containing electron dense particles and signet ring cells have usually one large vacuole which is electron dense lined and may contain electron dense particles. Developmental stages of these cell types show involvement of endoplasmic reticulum and Golgi bodies in granule formation. After glutaraldehyde fixation alone the only extremely electron dense components are particles in the compartment cells and signet ring cells implicating these as sites of vanadium localization, although not excluding other cell types.