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Scanning electron microscopy (SEM), especially low-voltage (1 KeV) high-resolution SEM, can be used in conjunction with stereo pair high-voltage (1 MeV) transmission electron microscopy (HVEM) of whole spread cells or thick sections effectively to correlate surface structure with internal structure. Surface features such as microvilli, pits, pseudopodia, ruffles, attached virus, and other surface-related morphologic characteristics can be identified using SEM, while underlying cytoskeletal structure and organelle organization can be viewed by HVEM of the same preparation. However, the need to "prepare" cells for electron microscopy precludes observation in the living state. The use of several types of video-enhanced light microscopy (VLM) permits observation of living cells such that certain surface and internal features can be observed at a relatively high level of resolution or detection. Thus, changes in living cells can be followed, and at appropriate times the cells may be chemically fixed or rapidly frozen and prepared for ultrastructural examination by electron microscopy. We have utilized VLM in conjunction with SEM and HVEM to correlate changes in shape and surface structure with changes in the internal structure of platelets. In addition, we have found it advantageous to use colloidal gold-labeling procedures, because these markers are detectable by all three forms of microscopy. Using this approach we have labeled platelet membrane GPIIb/IIIa, a receptor for RGD-containing adhesive proteins, with gold-fibrinogen or gold-anti-IIb/IIIa. The initial binding and subsequent movement of gold-fibrinogen-IIb/IIIa complexes in living platelets was followed by VLM. The movement of individual labels could be mapped. Subsequent observation by low-voltage (1 KeV) high-resolution SEM and HVEM permits visualization of the same individual receptors tracked by LM. The final position on the membrane or the position-in-transit when fixative was added was determined relative to surface ultrastructure (SEM) and internal, particularly cytoskeletal, ultrastructure (HVEM).
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We report uncultured rod-shaped magnetotactic bacteria from natural waters that biomineralize magnetic crystals in two different size ranges. Electron spectroscopic imaging of whole bacteria deposited over formvar-coated grids permitted a better visualization and measurement of the magnetosomes. All magnetosomes of individual bacteria could be observed by this technique. The magnetosomes formed one large chain, composed of three to four columns of crystals, disposed in parallel to the main axis of the bacteria. The magnetosomes ranged from 19 to 136 nm in length and 14 to 112 nm width. Smaller magnetosomes (less than 80 nm in length) localized mostly in extremities of the bacterial body while larger preferentially localized in the middle part of the cell. Electron spectroscopic diffraction and X-ray microanalysis indicate that both types of magnetosomes contain magnetite (Fe3O4). In projection, most magnetosomes seem to present pseudo-hexagonal morphologies described for magnetite. As the aspect ratios for smaller and larger magnetosomes are different, we suggest that different levels of control on biomineralization of magnetosomes may exist.
A 14-year-old girl demonstrated increased iron concentration and transferrin saturation, suggesting iron overload of unknown origin. Liver biopsy showed no fibrosis or hepatocytic atrophia. Nevertheless, Prussian blue reaction for histochemical detection of iron demonstrated very weak positive granules in a few hepatocytes on the periphery of hepatic lobules in close connection to bile capillaries. This very early stage of hemochromatosis was confirmed by TEM and EELS for iron accumulation inside hepatocytic lysosomes and residual bodies. Such siderosomes were scarce in number and iron content, compared to a case of manifested hemochromatosis and liver cirrhosis (Jonas L, Fulda G, Salemeh T, et al. Ultrastruct Pathol. 2001; 25: 111-118.). Liver iron concentration as measured by inductively coupled plasma-mass spectrometry (ICP-MS) and atomic absorption spectrometry (AAS) yielded 2.005 mg/g tissue dry weight, which was considered not significantly increased. In the absence of known causes for secondary iron overload, the early diagnosis was evidenced by genotyping, revealed homozygosity for the HFE gene C282Y mutation, demonstrating the presence of hereditary hemochromatosis.
The contractile responses to barium and the ultrastructure and ionic composition of mitochondria were studied in vascular smooth muscle. In normal rabbit portal anterior mesenteric vein (PAMV) and main pulmonary artery (MPA) smooth muscle mitochondria were frequently associated with the surface vesicles. The average distance between the outer mitochondrial and inner surface vesicle membrane was 4-5 nm. Ba contractures of MPA were tonic and of PAMV were phasic. Incubation of MPA and PAMV with Ba resulted in the accumulation of mitochondrial granules, followed in the MPA by massive mitochondrial swelling. Oligomycin and anoxia inhibited the appearance of mitochondrial electron-opaque granules and prevented the Ba-induced mitochondrial swelling in the MPA. Electron probe analysis of mitochondria in PAMV incubated with Ba and containing granules showed characteristic Ba signals over the mitochondria. Electron probe X-ray microanalysis also showed a highly significant (P < 0.001) correlation of P with mitochondrial Ba, in an estimated elemental ratio of approximately 3 Ba/4 P. Mitochondrial granules were still prominent after block staining of the osmium-fixed, Ba-loaded PAMV, but electron probe microanalysis showed no Ba, but only U, emissions. Tissues incubated with strontium had electron-opaque mitochondrial granules and deposits in the sarcoplasmic reticulum. X-ray microanalysis of mitochondria containing granules showed the presence of characteristic Sr and Ca emissions. The presence of Sr was similarly verified in the sarcoplasmic reticulum. These findings indicate the energy dependent uptake of divalent cations, in association with phosphate, by mitochondria in vascular smooth muscle in situ and the possibility that mitochondria may contribute to the regulation of intracellular divalent cation levels in smooth muscle.
This study was performed to determine the characteristics of esophageal dysplasia by scanning electron microscopy. A total of 82 esophageal biopsy specimens were taken from 30 patients who were divided into three groups. Group 1 patients had no known esophageal disease. Group 2 patients had squamous cell cancer. Group 3 patients had esophagitis. Mucosal biopsy specimens that had been diagnosed by light microscopy as normal, esophagitis, or dysplastic mucosa were examined by scanning electron microscopy. A characteristic appearance for each type of mucosa was recognized by scanning electron microscopy. A quantitative analysis of the scanning electron microscopy feature of microridge density showed a statistically significant difference not only between normal and dysplastic mucosa, but also between esophagitis and dysplastic mucosa. The addition of scanning electron microscopy to light microscopy may prove helpful in the diagnosis of dysplasia as well as in the understanding of the biologic behavior of dysplastic cells and possibly their relationship to esophageal carcinoma.
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The efficacy of conventional high vacuum scanning electron microscopy (SEM), environmental SEM (ESEM), and confocal laser scanning microscopy techniques in the assessment of cell-material interactions is compared. Specific attention is given to the application of these techniques in the assessment of the early morphological response of human osteoblast-like cells cultured on titanium dioxide. The processing of cells cultured for conventional high vacuum SEM leads to the loss of morphological features that are retained when using ESEM. The use of cytoskeletal labeling, viewed with confocal laser scanning microscopy, in conjunction with ESEM gives an indication of the changes to cell morphology as a consequence of incubation time in response to interactions at the biological/material interface.
Using electron microscope autoradiography, a study was made of the ultrastructure of early stages of muscle differentiation and 3H-thymidine (3H-T) labelled cells in the wall of the developing lymph heart of larvae of Rana temporaria L. The mononucleated postmitotic myoblasts with small bundles of thin and thick myofilaments deprived of Z-bodies were found in the lymph heart wall. No thin or intermediate-sized subsarcolemmal filaments were detected in the cytoplasm of these myoblasts. Myosatellites occurred under the basal lamina of muscle cells at stages 41-42. The primitive muscle-nerve junction was found at stages 44-45. Four hours after a single 3H-T administration only mononuclear cells without myofilaments were labelled. If the fixation was made 72 hours after a single 3H-T administration, the label was found, in addition, on the muscle cell nuclei. These data evidence that at the early stages of muscle differentiation in the developing lymph heart wall DNA synthesis and muscle specific protein synthesis are incompatible.
This report presents the interesting case of a 50-year-old white man with an unusual benign tumor composed predominantly of a proliferation of atypical endothelial cells combined with a variable inflammatory response. This case represents an instance of the recently renamed entity "histiocytoid hemangioma" in which two organ systems are involved. Both skin and bone showed typical lesions. No physical connection jointed the separate lesions. The results of examination by light microscopy, electron microscopy, and immunoperoxidase examination for lysozyme and Factor VIII are reported. The significance of this case is that it supports the concept of classifying similar vascular lesions, despite varied organ system origin, into a single entity, the histiocytoid hemangioma.
This report is the first description of the ultrastructure of the glandular elements in a so-called glandular Schwannoma (malignant peripheral nerve sheath tumor with glandular elements). The appearance of this uncommon tumor was typical by light microscopy. Electron microscopy of the sarcomatous component revealed features consistent with Schwann cell origin. Ultrastructure of the glands revealed a well-differentiated epithelium with basement membrane, desmosomes and the presence of microvilli with core rootlets, glycocalyx, and intracytoplasmic R-bodies. No cilia or blepharoplasts were present, arguing against ependymal origin of these glands. The microvilli with core rootlets, the glycocalyx and the R-bodies indicate the glandular epithelium was of the "intestinal type." The basis for the presence of such epithelium in Schwann cell neoplasms is uncertain, but pathologic induction-interaction between neural crest and mesenchyme may be the most plausible explanation.
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