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At least 1,171 records · Page 65Linked to original sources
PubMed2004

Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy.

The microstructural basis for the mechanical properties of blood vessels has not been directly determined because of the lack of a nondestructive method that yields a three-dimensional view of these vascular wall constituents. Here, we demonstrate that multiphoton microscopy can be used to visualize the microstructural basis of blood vessel mechanical properties, by combining mechanical testing (distension) of excised porcine coronary arteries with simultaneous two-photon excited fluorescence and second-harmonic generation microscopy. Our results show that second-harmonic generation signals derived from collagen can be spectrally isolated from elastin and smooth muscle cell two-photon fluorescence. Two-photon fluorescence signals can be further characterized by emission maxima at 495 nm and 520 nm, corresponding to elastin and cellular contributions, respectively. Two-dimensional reconstructions of spectrally fused images permit high-resolution visualization of collagen and elastin fibrils and smooth muscle cells from intima to adventitia. These structural features are confirmed by coregistration of multiphoton microscopy images with conventional histology. Significant changes in mean fibril thickness and overall wall dimension were observed when comparing no load (zero transmural pressure) and zero-stress conditions to 30 and 180 mmHg distension pressures. Overall, these data suggest that multiphoton microscopy is a highly sensitive and promising technique for studying the morphometric properties of the microstructure of the blood vessel wall.

Animals↗
PubMed2006

Nonperturbative chemical imaging of organelle transport in living cells with coherent anti-stokes Raman scattering microscopy.

Nonperturbative monitoring of intracellular organelle transport in unstained living cells was achieved with coherent anti-Stokes Raman scattering (CARS) microscopy. To avoid possible interference with the organelle transport introduced by laser radiation, we first examined different illumination conditions. Using a new photodamage criterion based on morphological changes of the cells, we determined the threshold values of both pulse energy and average power at relevant wavelengths. Under excitation conditions much milder than the threshold levels, we were able to monitor the motions of lipid droplet (LD) organelles in steroidogenic mouse adrenal cortical (Y-1) cells with CARS microscopy in real time without perturbations to the cells. Particle tracking analyses revealed subdiffusion as well as active transport of LDs along microtubules. Interestingly, LD active transport is only present in Y-1 cells that rounded up in culture, a morphological change associated with steroidogenesis, suggesting possible involvements of LD active transport in the latter. Simultaneous imaging of LDs and mitochondria with CARS and two-photon fluorescence microscopy clearly showed that interactions between the two organelles could be facilitated by high LD motility. These observations demonstrate CARS microscopy as a powerful noninvasive imaging tool for studying dynamic processes in living cells.

Animals↗
PubMed2006

Maxillary sinus augmentation using an engineered porous hydroxyapatite: a clinical, histological, and transmission electron microscopy study in man.

Porous hydroxyapatite (HA) is a calcium-phosphate-based material that is biocompatible, nonimmunological, and osteoconductive, and has a macroporosity of about 200 to 800 microm. The pores seem to be able to induce migration, adhesion, and proliferation of osteoblasts inside the pore network and to promote angiogenesis inside the pore system. The aim of this study was to evaluate the clinical behavior and the histological and ultrastructural aspects of porous HA in maxillary sinus augmentation procedures. Twenty-four patients (19 men, 5 women; average age 53.4 years) in good general physical and mental health and with partially or completely edentulous maxillae were selected for this study. Six months after sinus floor elevation, at the time of dental implant placement, biopsies were carried out under local anesthesia. These bone cores were cut in half and were processed for light and transmission electron microscopy. After a mean 3 years after implantation, all implants are clinically in function and no surgical or prosthetic complications have occurred. Under light microscopy, newly formed bone was 38.5% +/- 4.5%, whereas the residual biomaterial represented 12% +/- 2.3% and the marrow spaces represented 44.6% +/- 4.2%. In addition, in the majority of cases, the biomaterial particles were in close contact with the bone, which appeared compact with the characteristic features of well-organized lamellar bone. A cement-like line was slightly visible at the bone-biomaterial interface, but there were no gaps or interposed connective tissue in between. A high quantity (about 40%) of newly formed bone was present. Bone was closely apposed to the biomaterials particles as shown in light microscopy and transmission electron microscopy. Moreover, no signs of inflammatory cell infiltrate or foreign body reaction were present. Also, most of the biomaterial was resorbed and only a small quantity (a little more than 10%) was still present. The results of our study show that porous HA can be a suitable synthetic material for bone regeneration in maxillary sinus augmentation procedures.

Absorbable Implants↗
PubMed1985

Rapid identification of periodontal pathogens in subgingival dental plaque. Comparison of indirect immunofluorescence microscopy with bacterial culture for detection of Bacteroides gingivalis.

A large body of research implicates Bacteroides gingivalis in the etiology of adult periodontitis, however, the application of this information to clinical diagnosis and treatment has been hampered by the need for a simple, rapid, and reliable means of detecting this microorganism. In the present study, indirect immunofluorescence microscopy using species specific, polyclonal antisera and a monoclonal antibody was evaluated in the clinical identification and quantitation of B. gingivalis in human subgingival dental plaque. One hundred and twenty subgingival plaque samples were obtained from predetermined sites by means of sterile paper points from 20 human subjects including 10 adult periodontitis patients and 10 periodontally normal subjects. The proportions of cultivable B. gingivalis in each sample were determined following anaerobic culture on nonselective blood agar media and selective media containing kanamycin. These results were then compared to quantitative estimates of B. gingivalis by indirect immunofluorescent microscopic evaluation of heat-fixed plaque smears. Using both immunofluorescence microscopy and bacterial culture, the present study confirms the importance of B. gingivalis in adult periodontitis previously described by culture. The organism was cultivable from 70% of the adult periodontitis patients but not from any of the normal adults. In contrast, indirect immunofluorescence microscopy detected the organism in up to 40% of the subgingival sites in 100% subgingival sites in 100% of the adult periodontitis patients as well as four sites in the periodontally normal subjects. The sensitivity of indirect immunofluorescence microscopy compared to culture ranged from 91 to 100% while the specificity varied from 87 to 89%.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1999

Evaluation of initial attachment of human gingival fibroblast cells to biodegradable membranes in vitro by light and scanning electron microscopy.

Guided tissue regeneration procedures using resorbable membranes have become accepted therapy for treating periodontal defects. Resorbable collagen and synthetic polylactide and polyglycolide copolymer membranes have been found to support regeneration and preclude the need for surgical removal. This study was undertaken to assess and compare the initial attachment of human gingival fibroblast cells to four collagen-based membranes (fascia lata, fascia temporalis, dura mater, and Type I bovine collagen) and a synthetic polylactic acid-based membrane (resolut). Human gingival fibroblasts were grown from explants of normal tissue obtained during surgical reduction of retromolar tissues. Membrane specimens were placed in separate culture wells and incubated with fibroblasts for one hour. The number of adherent cells was evaluated by light microscopy using an ocular grid system and detailed examination was performed by scanning electron microscopy. The results of evaluation by light microscopy indicated that initial cell attachment was significantly less in the polylactic acid-based membrane group than in the collagen-based membrane groups (P < 0.01). However, no significant differences were found among the collagen membrane groups in terms of fibroblast attachment (P > 0.01). Scanning electron microscopy examination of fibroblasts cultured directly on barrier membranes indicated that the collagen-based membranes appeared to facilitate cell attachment, whereas the polylactic acid-based membrane exhibited a morphology that was not conducive to attachment of human gingival fibroblasts. Based on these limited in vitro results, it appears that collagen-based membranes offer greater potential than polylactic acid-based membranes for guided tissue regeneration at surgical sites.

Absorbable Implants↗
PubMed1980

Freeze microscopy of the endometrium in ectopic pregnancy.

A series of 60 patients with suspected ectopic pregnancy was reviewed in order to assess the diagnostic value of freeze microscopy of endometrial curettings. By this procedure the correct diagnosis has been make in 82 per cent of patients, i.e. it established whether the patient was pregnant and whether the pregnancy was intra- or extrauterine. Eighteen patients had an ectopic pregnancy. 13 of them were detected primarily on microscopy of frozen sections, while the remaining 5 were erroneously classified as non-pregnant. The diagnostic sensitivity of freeze microscopy proved to be equal to microscopy of the subsequent paraffin sections. A diagnostic and therapeutic schedule can be set up for patients with suspected ectopic pregnancy when the histological appearances of the frozen sections and the result of the HCG reaction are known.

Endometrium↗
PubMed1995

Confocal laser scanning microscopy of small-intestinal mucosa in celiac disease.

BACKGROUND: Light microscopy of jejunal biopsy specimens is routinely used to diagnose celiac disease. Confocal laser scanning microscopy offers research advantages, since thin optical sectioning can be performed without mechanical damage to the specimen. Fixed as well as non-fixed specimens can be studied. METHODS: Confocal laser scanning microscopy was used to study the stanining of wheat germ agglutinin, which binds to glycoconjugates of the enterocyte, and rhodamine phalloidin, which binds to the F-actin of the cells. RESULTS: In healthy mucosa the wheat germ agglutinin labeling showed a strong and punctate staining of microvilli, which outlined a convoluted surface. Phalloidin-labeled actin formed a three-dimensional cage at the cell membrane. In the crypt-hyperplastic mucosa, both staining patterns were irregular, and the cytoskeleton was disorganized. CONCLUSIONS: Confocal laser scanning microscopy offers the possibility to study the distribution of surface and cytoskeleton markers in thick, structurally intact specimens.

Celiac Disease↗
PubMed1987

Electron microscopy in the diagnosis of percutaneous fine needle aspiration specimens.

Five years experience in the application of electron microscopy to fine needle aspiration biopsy specimens is reviewed. In an initial evaluation, 200 consecutive unselected specimens were examined; 89 proved diagnostic and, in a third of these, electron microscopy gave additional information that was often essential to diagnosis. Negative specimens resulted almost entirely from failure to obtain an adequate amount of material. Results were improved by the adoption of a preparation technique involving concentration of the cells in bovine serum albumin and by the inspection of a rapidly stained smear at the time of the aspiration procedure, with a further needle pass for electron microscopy being performed if necessary. Despite the small size of specimens, adequate examination was usually possible and electron microscopy has proved of value in the diagnosis of tumor samples acquired by fine needle aspiration in the same way as has been established with the larger sized specimens obtained by conventional biopsy and surgical resection.

Adenocarcinoma↗
PubMedDate not supplied

Microcomputer application: an image database system for electron microscopy.

An electron microscopist in a small electron microscopy laboratory that processes 200 cases per year can generate at least 2000 electron micrographs within this time period. These electron micrographs are stored in filing cabinets, which slowly encroach on existing laboratory space. Related image data are often located in separate filing systems. This type of storage system is inefficient for rapid retrieval of sets of electron micrographs for research, teaching, or retrospective review of patient material. An electron microscopy image base, which is implemented on a microcomputer, can provide flexible and simultaneous access to both digitized electron micrographs and their relevant textual data. In addition to the advantage of more efficient retrieval, electronic storage of textual data and digitized electron micrographs also offers the advantage of decreased storage space for this type of data. Implementation of the initial version of the electron microscopy image base demonstrated that future versions must provide improved resolution of the digitized electron micrograph and sufficient electronic mass storage to support such a large image base. Ongoing developments in database technology and mass storage make it worthwhile to pursue additional refinements of the electron microscopy image base.

Database Management Systems↗
PubMedDate not supplied

Electron microscopy in the diagnosis of small round cell tumors of bone.

Small round cell tumors involving bone can present problems in differential diagnosis by light microscopy. In exploring the role of electron microscopy in this situation, seven small cell osteosarcomas and seven mesenchymal chondrosarcomas were examined by electron microscopy and compared with typical and atypical Ewing's sarcomas. There is much overlap in the ultrastructural features of these tumors, but electron microscopy is helpful to establish or confirm a diagnosis of typical Ewing's sarcoma and, if representative matrix is present, of small cell osteosarcoma.

Bone Neoplasms↗
PubMedDate not supplied

Usefulness of electron microscopy in the diagnosis of "small" round cell tumors of the sinonasal region.

The sinonasal region is known to harbor several types of tumors that belong to the general category of "small" round cell tumors and offer considerable diagnostic challenges. This study evaluated 33 cases of such tumors by electron microscopy to characterize their ultrastructural features in conjunction with immunohistochemistry, in an attempt to define diagnostic criteria of various types. Electron microscopy was useful in the proper classification of tumors in 27 cases: esthesioneuroblastoma (EN), 12; undifferentiated carcinoma, 6; melanoma, 3; lymphoma, 3; melanotic neuroectodermal tumor, 1; rhabdomyosarcoma, 1; and pituitary adenoma, 1. In the remaining six cases, the ultrastructural features were those of poorly differentiated carcinomas. They usually exhibited some epithelial characteristics as well as neuroendocrine features by immunohistochemistry and electron microscopy. These tumors could be best described as poorly differentiated neuro-endocrine carcinomas (malignant neuroepitheliomas). The most controversial diagnostic problems existed between the tumors categorized as esthesioneuroblastomas and neuroendocrine (NE) carcinomas. Esthesioneuroblastomas were characterized by uniform round nucleated cells with variable amounts of dendritic processes containing numerous dense core granules ranging from 150 to 350 nm in the perikarya and dendritic processes. Dendritic processes contained longitudinally arranged neural tubules and revealed an occasional synaptic junction. In three of the 12 cases of EN, cells with the appearance of sustentacular cells were recognized by electron microscopy. The NE carcinomas usually consisted of closely packed round cells with scanty cytoplasm that lacked any feature of neuroblastic cells. The tumor cells in this category often were epithelioid in appearance and exhibited a varying degree of cytokeratin positivity. Neuron-specific enolase was also positive in all cases, further suggesting their neuroepithelial nature. The greatest difference between EN and NE carcinomas was the absence of sustentacular cells in NE carcinomas. Immunohistochemical and electron microscopic studies are essential in the differential diagnosis of EN and NE carcinomas, because their microscopic appearance is very similar. The study indicates that EM is useful in the diagnostic categorization of sinonasal tumors of uncertain nature, particularly when it is used in conjunction with immunohistochemistry.

Carcinoma↗
PubMedDate not supplied

Immunoelectron microscopy for chromogranin A in small cell neuroendocrine carcinoma of lung.

To see if immunoelectron microscopy can improve localization of neurosecretory granules, postembedding immunolabelling for chromogranin A was performed on 15 examples of small cell anaplastic (neuroendocrine) carcinomas primary in lung, five cases of bronchopulmonary carcinoids, and two cases of pheochromcytoma; both the carcinoids and pheochromocytomas had neurosecretory granule-rich cytoplasm by routine electron microscopy and some neurosecretory granules were identified in each of the small cell carcinomas. Immunolabeling for chromogranin A resulted in many colloidal gold particles over cytoplasmic secretory granules in both pheochromcytomas and four of the carcinoids. One carcinoid that was focally positive by immunoperoxidase staining was negative by immunoelectron microscopy. None of the 15 cases of small cell carcinoma stained for chromogranin A using immunoperoxidase techniques, but three had a small number of secretory granules weakly labeled by the anti-chromogranin A/colloidal gold complex. Immunoelectron microscopy, at least using standard glutaraldehyde fixation and epoxy resin embedding, does not increase the sensitivity of neurosecretory granule identification in small cell neuroendocrine carcinomas of the lung. Results of other studies of this neoplasm suggest that despite transcription of chromogranin genes, synthesis of the specific protein may occur to a limited extent or not at all.

Carcinoid Tumor↗
PubMed1998

In vivo tandem scanning confocal microscopy in acanthamoeba keratitis.

The in vivo confocal microscopy technique provides us with a real-time, non-invasive way of examining the human cornea. The most important advantage of this type of microscopy is to reveal the etiologic agents in infectious keratitis such as Acanthamoeba keratitis. We present several representative cases of Acanthamoeba keratitis, which were diagnosed in their early stages using in vivo confocal microscopy and managed based on that diagnosis. In our Acanthamoeba keratitis cases, highly-reflective round or ovoid organisms with a diameter of about 10-25 um were visualized distinctly against relatively-dark normal parenchymal structures, such as epithelial cells or keratocyte nuclei. Double-walled structures of Acanthamoeba cysts were clearly demonstrated in some cases. We can confirm that in vivo tandem scanning confocal microscopy is a powerful diagnostic tool for identifying the infecting organisms in Acanthamoeba keratitis.

Acanthamoeba↗
PubMedDate not supplied

Confocal microscopy of stage 4 diffuse lamellar keratitis with spontaneous resolution.

PURPOSE: To report confocal microscopic findings at the onset of stage 4 diffuse lamellar keratitis and after its resolution. Stage 4 is the most severe form of diffuse lamellar keratitis. Its incidence is approximately 1 in 5000 and is associated with stromal melting, deep flap folds, central haze, hyperopic shift, irregular astigmatism, and severe decrease in visual acuity. METHODS: A 22-year-old woman underwent bilateral uncomplicated laser in situ keratomileusis (LASIK) for myopia. Postoperative course in the right eye was uneventful; however, in the left eye, stage 4 diffuse lamellar keratitis developed. Confocal microscopy examination was performed in both eyes at the onset of the syndrome and after its resolution. Findings in the eye with diffuse lamellar keratitis (left eye) were compared with the uninvolved (right) eye. RESULTS: The condition improved spontaneously and 2 years later, slit-lamp microscopy showed resolution of the folds and haze with subsequent improvement of visual acuity. However, confocal microscopic examination in the left eye revealed a persistent stromal subclinical haze on both sides of the lamellar cut and prominent folds that extended into the anterior stroma. CONCLUSION: Confocal microscopy revealed that in spite of normal appearance on slit-lamp microscopy, micromorphological alterations persisted after spontaneous resolution of stage 4 diffuse lamellar keratitis.

Adult↗
PubMed1992

Localization of a 230-kD parasitophorous vacuole membrane antigen of Plasmodium berghei exoerythrocytic schizonts (LSA-2) by immunoelectron and confocal laser scanning microscopy.

Using antiserum to a 230-kD parasitophorous vacuole membrane (PVM) antigen of Plasmodium berghei exoerythrocytic schizonts as a specific probe for the PVM, we studied the three-dimensional structure of this membrane within infected host cells by immunoelectron microscopy and confocal laser scanning microscopy at 3, 4, and 50 hr after sporozoite invasion. Fluorescent label was not detected at 3 hr, but was associated with the cytoplasm of 24-hr-old exoerythrocytic parasites. Specific labeling of the PVM was not observed by immunoelectron microscopy until 50 hr, when numerous vesicles and finger-like projections of the PVM were found in the cytoplasm of infected host cells. Labeled vesicles were often isolated and located at the periphery of the infected hepatocyte. Confocal microscopy demonstrated that these vesicles formed discontinuous chains that extended from 3-10 microns away from the parasite. These structures appear to be similar to the membranous clefts of Plasmodium-infected erythrocytes, and may be important in the movement of host or parasite proteins within infected hepatocytes.

Animals↗
PubMed1998

[Fluorescence microscopy for detection of frauds in ground coffee].

Currently, frauds in ground coffee are detected with traditional microscopy, which is a tedious and not very precise method. Therefore, a more efficient and rapid microscopical method based on fluorescence, was proposed. For the microscopy analysis, pure coffee was compared to coffee samples adulterated with rye, barley, corn and wheat at 2.5, 5.0, and 10.0% levels. Starch granules were detected and identified immediately with fluorescence microscopy, in the adulterated coffee mixtures, quite different from optical microscopy where no frauds were detected with certainty. The suggested fluorescence method for the detection of cereal frauds in coffee was found to be more efficient and precise than the current official one.

Coffee↗
PubMed1998

[Fetal hypotrophy: morphological pictures of placenta in light and electron microscopy of female residents of the Czenstochow region].

The 16 afterbirths samples obtained from pregnancies complicated by idiopathic fetal hypotrophy was examined. The criteria of hypotrophy were established: the duration of gestation above 36 weeks, the birth-weight was less than 2500 g. The control group included 8 afterbirth samples from physiological full-term pregnancies. It was performed macroscopic examination, light microscopy and transmission electron microscopy. Macroscopically, no changes were found in the study group apart from the small differences of the weight. Light microscopy showed the retardation of placenta maturation from fetus hypotrophy group, congestion of the villi and presence of the numerous calcifications. Transmission electron microscopy showed the changes of placental barrier elements: adaptation processes and chronic progressive damage of the cells (exfoliation of the endothelial cells from the basal membrane).

Adult↗
PubMed1999

Maximization of skin capillaries during intravital video-microscopy in essential hypertension: comparison between venous congestion, reactive hyperaemia and core heat load tests.

Intravital capillary video-microscopy is a dynamic method for studying skin capillaries. The technique of direct intravital microscopy (without dyes) depends on the presence of red blood cells inside capillaries for their identification. The aim of the present study was to compare different techniques to try to establish the best method for maximizing the number of visible perfused capillaries during intravital capillary microscopy. We compared the effects of venous congestion with those of post-occlusive reactive hyperaemia (Study 1). We also investigated venous congestion followed first by post-occlusive reactive hyperaemia and then by a core heat load test (Study 2). Finally we investigated venous congestion followed by post-occlusive reactive hyperaemia combined with venous congestion (Study 3). In Study 1, capillary density increased with venous congestion from a baseline value of 74+/-2 (mean+/-S.E.M.) per field to 82+/-3 per field (P<0.0001; analysis of variance). With reactive hyperaemia, there was an apparent decrease in visible capillary density to 69+/-2 per field. In Study 2, baseline capillary density was 69+/-4 per field, and this increased significantly with venous congestion to 74+/-4 per field (P=0.01). With both reactive hyperaemia and core heat load, the apparent density was 62+/-4 per field. In Study 3 the baseline density was 70+/-2 per field, and this increased significantly with venous congestion to 80+/-3 per field (P<0.0001). With reactive hyperaemia combined with venous congestion, the density was 81+/-3 per field (P=0.328 compared with venous congestion alone). The results show that venous congestion at 60 mmHg for 2 min is the most effective method for visualization of the maximal number of perfused skin capillaries during intravital video-microscopy.

Adult↗