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Results for “MICROSCOPY, PHASE CONTRAST”

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PubMed1992

[Changes in the form and ultrastructure of the smooth-muscle cells in the human aorta in prenatal ontogeny].

The shape of smooth muscle cells (SMC) was analysed using the phase contrast microscopy of cell suspensions obtained by alcohol-alkali dissociation, as well as the semithin sections prepared in perpendicular planes. The phenotype of SMC was analysed using transmission electron microscopy. The shape of SMC changes from preferentially round to preferentially spindle-like and stellate one during development. The differentiation of SMC is accompanied with the increase in the contractile apparatus content and in the decrease in the content of synthetic organelles.

Actin Cytoskeleton↗

PubMed1989

[Microscopic hematuria. Semiologic value in urology. Management of microscopic hematuria].

Microscopic hematuria, a frequent cause for consultation, poses a problem regarding its significance. It is essential that a curable pathology is not neglected. In the absence of associated clinical data, nephrological orientation of investigations depends on the study of urinary sediments and phase contrast microscopy. Where there is a urological orientation, half the lesions involve the lower urinary tract; the use of flexible urethrocystoscopy has been a true advance in terms of its simplicity and reliability. When a lesion is demonstrated the question still remains as to whether it is in fact responsible for the microscopic hematuria. It is best to remain prudent when a poorly significant lesion is discovered. Since 20% of microscopic hematurias remain unexplained, how far should one go in the paraclinical investigation and follow up of involved patients?

Adult↗

PubMed1980

Kinetics of ADP-induced human platelet shape change: apparent positive cooperativity.

The kinetics of ADP-induced human platelet shape change have been examined. Initial velocities of platelet shape change were estimated by two methods: (1) the slope of the initial decrease in light transmission through stirred, citrated platelet-rich plasma, and (2) direct examination of platelet morphologies by phase-contrast microscopy. In both cases, a value of the Hill coefficient, NH, significantly greater than 1 is obtained (2.0 +/- 0.2 and 1.8 +/- 0.2, respectively). The observed elevated value of NH is not due to a substantial fraction of the ADP being platelet bound, the presence of factors in the plasma, platelet heterogeneity, or the influence of the rate of platelet shape change reversion. Our observations suggest that ADP-induced platelet shape change may be a positively cooperative or "threshold" type response.

Adenosine Diphosphate↗

PubMed1975

Neurons from fetal rat brain in a new cell culture system: a multidisciplinary analysis.

A new culture system for cells from the mammalian brain was developed by a modification of a previously established technique. This modification involved the use of fluorodeoxyuridine and adult horse serum. The cultures contained large, easily visualized neurons both isolated from other neurons and in networks of varying complexity. These cells were large enough to permit reliable intracellular electrophysiologic recording and were often sufficiently dispersed to allow examination of membrane responses to iontophoretically applied neurotransmitter candidates. Many responses characteristic of central neurons in situ were seen, including evoked and spontaneous action potentials, complex patterns of inhibitory and excitatory post-synaptic potentials, and neurotransmitter-induced membrane responses. These preparations were examined by phase contrast microscopy, by light microscopy after silver impregnation and by Nomarski interference optics. Total choline acetyltransferase (CAT) activity was little changed and specific activity was increased in the new culture system as compared with the earilier system. Conditions which gave the highest specific activity of CAT also provided the best cultures from the standpoint of electrophysiologic and morphologic analysis. This new approach will allow, in culture, detailed multidisciplinary analyses of individual neurons and small networks of neurons from the mammalian brain.

Acetylcholine↗

PubMed1995

A threshold decrease for electrically stimulated motor responses of isolated aging outer hair cells from the pigmented guinea pig.

When outer hair cells are isolated from guinea pig cochleas and are placed in normal Hank's medium, they exhibit aging as a slow tonic reduction in length and increase in diameter. During this time the lateral subsurface cisternae become progressively vesiculated and the optical density of the border seen under phase-contrast microscopy decreases. A study of 65 outer hair cells was carried out using video imaging of this process. The base of each cell bonded to the Petri dish and the motility of the cuticular plate was recorded in two ways. To quantify the slow contraction of each preparation, the dimensions of the cell were measured from video replay. Displacements of the cuticular plate in response to an alternating electric field in line with the cell axis were also monitored using a video tracking technique. The amplitude of a 1 Hz stimulus required to cause a visually detectable motor response above baseline noise decreased as the cell degraded. Typically, fresh cylindrical cells exhibiting high optical contrast showed relatively small movements for field strengths up to 2 kVm-1. However, as the cell depolarized, the rigidity initially decreased and the cell could respond to field strengths down to 0.1 kVm-1 before cell death ultimately occurred. Such a threshold phenomenon in the isolated OHC has not been demonstrated directly until now. This result explains the variability of electromotility in aging in vitro preparations from the cochlea.

Animals↗

PubMed1989

Distinction between vascular smooth muscle cells and myoepithelial cells in primary monolayer cultures of human breast tissue.

We report on the discrimination of vascular smooth muscle cells and myoepithelial cells in primary cultures of human breast tissue. Breast tissue was disaggregated enzymatically and the resulting organoids seeded in monolayer culture on collagen-coated plastic in serum-free medium CDM3a. Two main types of organoids were present after enzymatic digestion. One resembled small blood vessels and the other interlobular ducts or acini of the breast gland epithelium. Within 3 to 8 d after plating the organoids migrated into typical monolayer islets. These monolayer islets were evaluated using phase contrast microscopy and further tagged with monoclonal antibodies for immunocytochemical demonstration of Factor VIII-related antigen, muscle iso-forms of actin, type IV collagen, vimentin, desmin, and keratins. It is concluded that vascular smooth muscle cells resembled myoepithelial cells by expressing vimentin filaments, depositing type IV collagen, and showing immunoreactivity to muscle iso-forms of actin. However, whereas vascular smooth muscle cells were associated with endothelial cells and sometimes expressed desmin, myoepithelial cells appeared together with luminal epithelial cells and expressed cytokeratins.

Actins↗

PubMed1984

Lobar deposition of fibers in the rat.

Nineteen pathogen free male rats were exposed to a ceramic fiber dust cloud with an average of 709 f/mL for 5 days, 6 hours per day, by nose only, to study the deposition of fibers in the different lung lobes. The count median diameter and length of the airborne fibers were 0.53 micron and 3.7 microns respectively. Five days after the last day of exposure, the rats were sacrificed; lungs were excised, and lobes were separated, weighed and digested. The digested lungs were filtered on membrane filters. These filters as well as samples of the airborne fiber cloud were evaluated by optical phase contrast microscopy at 1000 X total magnification. Fiber deposition was calculated from the concentration of airborne fibers, the minute volume, the duration of exposure and the number of fibers in the digested lung tissue for various fiber length and diameter categories. Fiber deposition in all the lobes varied over a narrow range, 5.43% for the right diaphragmatic lobe to 8.38% for the right apical lobe. The fiber burden for all lobes was found to be weight dependent. For the right apical lobe, the anatomical characteristics also influenced fiber deposition. Fiber size distributions in the various lobes were not significantly different.

Animals↗

PubMed1985

Interface analysis of titanium and zirconium bone implants.

A thin layer of titanium or zirconium was evaporated onto the surface of a plastic implant which was then inserted in the rabbit tibial metaphysis for six months. The implants and surrounding bone were cut out en bloc and sectioned for phase contrast microscopy, scanning- and transmission-electron microscopy. The intact bone-to-metal interface in the case of titanium revealed a fibrous tissue-free boundary zone with a 200-400 A thick proteoglycan coat immediately adjacent to the titanium oxide. Thereafter collagen filaments were seen and, at approximately 1000 A from the interface, collagen bundles. The tissues surrounding the zirconium-coated implant consisted of a 300-500 A thick proteoglycan layer, followed by a zone with collagen filaments and collagen bundles, not closer to the zirconium oxide than a few thousand A.

Animals↗

PubMed1996

The influence of periodontitis on the subgingival flora around implants in partially edentulous patients.

The hypothesis that teeth act as reservoirs of micro-organisms for the colonization of oral implants has recently been stated several times. The present study aimed at examining, in partially edentulous patients with severe periodontitis, whether pockets around teeth and implants harbored a comparable micro-flora. In 6 patients (3 with refractory periodontitis and 3 with advanced chronic adult periodontitis), plaque samples were taken from a deep and shallow pocket around both teeth and implants for differential phase contrast microscopy and DNA probe analysis. The results showed important differences in the sub-gingival flora between the 2 disease groups, as well as between deep and shallow pockets, around both implants and teeth. On the other hand, when pockets around teeth and implants with equal depths were compared a striking similarity was observed in the microbial composition. These observations confirm the hypothesis that pockets around teeth act as a reservoir and highlight the importance of periodontal health when oral implants are planned.

Adult↗

PubMed2003

Establishment and characterization of a human ovarian granulosa tumor cell line (HSOGT).

We successfully established a novel ovarian granulosa tumor cell line (HSOGT). The tumor tissue of the ovary was derived from a 25 year-old Japanese woman under her consent. The cell line was maintained for over 14 months, subcultured more than 73 times, and had a population doubling time of 18.9 hours. Phase contrast microscopy displayed a pavement-like arrangement without contact inhibition. The chromosome number showed a wide distribution of aneuploidy and the mode was 83; many marker chromosomes were observed. The HSOGT was also successfully xenotransplanted into nude mice. The cell line produced estradiol and has preserved some characters of granulosa cells with stable growth in vitro. We firmly believe that this cell line will be a most useful tool for endocrinological investigation of human granulosa cells.

Adult↗

PubMed1978

[Morphology and deformability of erythrocytes in muscular dystrophy].

In recent years, the presence of red cell morphological abnormalities in patients with Muscular Dystrophy has made the object of numerous, often contradictory reports. A possible source of such confusion may lie in the fact that human erythrocytes are extremely sensitive to morphologic transformations resulting from various manipulations or environmental conditions in vitro. We have examined the morphology and deformability of erythrocytes from 7 patients with Duchenne and 9 patients with Steinert (myotonic) Muscular Dystrophy. To avoid preparation artifacts, fresh, unwashed red cells suspended in their own plasma were examined under phase contrast microscopy for the presence of either echinocytes and stomatocytes. Deformability was measured by filtration of dilute cell suspensions at constant flow rate through nucleopore membranes (nominal pore diameter = 3 micron). No significant difference was found between the patients' cells and those of 22 healthy volunteer controls. We conclude that previously reported abnormalities may have been the result of preparation artifacts. It appears possible, however, that erythrocytes from Muscular Dystrophy patients may be more sensitive than normal ones to certain stimuli originating from red cell manipulations in vitro.

Adolescent↗

PubMed1998

Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development.

The formation of complex bacterial communities known as biofilms begins with the interaction of planktonic cells with a surface in response to appropriate environmental signals. We report the isolation and characterization of mutants of Pseudomonas aeruginosa PA14 defective in the initiation of biofilm formation on an abiotic surface, polyvinylchloride (PVC) plastic. These mutants are designated surface attachment defective (sad ). Two classes of sad mutants were analysed: (i) mutants defective in flagellar-mediated motility and (ii) mutants defective in biogenesis of the polar-localized type IV pili. We followed the development of the biofilm formed by the wild type over 8 h using phase-contrast microscopy. The wild-type strain first formed a monolayer of cells on the abiotic surface, followed by the appearance of microcolonies that were dispersed throughout the monolayer of cells. Using time-lapse microscopy, we present evidence that microcolonies form by aggregation of cells present in the monolayer. As observed with the wild type, strains with mutations in genes required for the synthesis of type IV pili formed a monolayer of cells on the PVC plastic. However, in contrast to the wild-type strain, the type IV pili mutants did not develop microcolonies over the course of the experiments, suggesting that these structures play an important role in microcolony formation. Very few cells of a non-motile strain (carrying a mutation in flgK) attached to PVC even after 8 h of incubation, suggesting a role for flagella and/or motility in the initial cell-to-surface interactions. The phenotype of these mutants thus allows us to initiate the dissection of the developmental pathway leading to biofilm formation.

Biofilms↗

PubMed1981

Effects of high hydrostatic pressure on normal and neoplastic rat cells in culture.

Four types of rat cells in culture were exposed to hydrostatic pressures in the range 1-1,500 bar. Each applied pressure was constant for half an hour. The morphological effects of pressure application were studied by phase contrast microscopy, and mortality was measured by total cell counts and the trypan blue exclusion test. Morphological changes characterized by cell rounding were observed in secondary fetal brain cells and fibroblasts at about 700 bar. In two permanent neoplastic neurogenic cell lines similar changes occurred at 1,000 to 1,100 bar. When approximately 50% of the cells were rounded, mortality began to increase, as compared with controls. This was gradual in secondary cells and comparatively abrupt in the permanent neoplastic lines. Malignant cells in culture may therefore be more resistant to hydrostatic pressure than their normal counterparts.

Animals↗

PubMed1962

Cytological observations on the chromatin bodies of two Bacillus species.

Chatterjee, B. R. (Baylor University College of Medicine, Houston, Tex.) and Robert P. Williams. Cytological observations on the chromatin bodies of two Bacillus species. J. Bacteriol. 83:1112-1118. 1962.-The nature of the chromatin bodies in Bacillus anthracis and B. megaterium has been studied intracellularly as well as after isolation from the bacilli. Preparations were examined by phase-contrast microscopy in the living state and by acid-Giemsa and Feulgen staining. In cells from young cultures, chromatin bodies occupy a continuous, long, central area in the bacteria. As the cells mature, the bodies lose their continuity and divide into two halves. In cells grown for 24 hr or longer, the chromatin bodies become more complex, and appear as multiple, spherical, interconnected constellations. Nuclear staining reveals that only the peripheral area, whether in cells from young or older cultures, takes up the stain, leaving a clear, central core. Discrete chromatin bodies are liberated from cells grown for longer periods after disintegration of the bacterial cell wall. These discrete bodies have been isolated from whole bacteria by treating washed, buffered saline suspensions of old cells with high concentrations of lysozyme followed by digestion with purified pancreatic lipase. The isolated chromatin bodies retain the same structure and appearance as inside the cells and show the same staining characteristics. Observations suggest that the chromatin bodies are discrete and circumscribed in nondividing, resting-phase bacteria.

Animals↗

PubMed1998

The effect of minoxidil on keratocyte proliferation in cell culture.

PURPOSE: To determine if minoxidil inhibits keratocyte proliferation in a nontoxic manner. METHODS: Rabbit keratocytes were cultured in Eagle's minimum essential medium supplemented with fetal bovine serum. Minoxidil varying in concentration from 10(0) to 10(3) micrograms/ml was added to the culture medium and incubated for 7 days. The cultures were inspected for morphologic appearance and the cell number was determined at 1, 3 and 7 days after the addition of minoxidil. After 7 days of incubation, minoxidil was withdrawn from the cell culture medium and the cells were examined 3 and 7 days thereafter. In addition, a nonradioactive cytotoxic assay was performed to determine if toxicity is associated with the presence of minoxidil. RESULTS: Minoxidil inhibited keratocyte proliferation in a dose-dependent fashion. 29% of control growth was achieved when keratocytes were cultured for 7 days in 10(3) micrograms/ml, whereas 82% control growth was achieved when keratocytes were cultured in 10(2) micrograms/ml of minoxidil. Intermediate concentrations between 10(2) and 10(3) micrograms/ml produced a linear decline in cell counts in a dose-dependent fashion. The concentration of minoxidil required for 50% control growth at 7 days extrapolated from the dose-response curve was 600 micrograms/ml. Upon withdrawal of minoxidil, cell counts returned to baseline for concentrations of 10(2) micrograms/ml or less. Phase contrast microscopy revealed that the presence of minoxidil was associated with intercellular separation, enlargement of cell bodies and elongated processes. After the withdrawal of minoxidil, the cells in all media reassumed the morphological features of normal keratocytes which included a regular fusiform shape and extensive intercellular contact. The nonradioactive cytotoxic assay revealed the lack of cytotoxicity at all concentrations of minoxidil based on a lack of lactate dehydrogenase release. CONCLUSIONS: Minoxidil inhibits keratocyte proliferation by a nontoxic mechanism. It might be particularly useful for modulating corneal wound healing following excimer laser photorefractive keratectomy.

Animals↗

PubMed1990

Microbial patterns in pooled subgingival plaque samples from young adults with advanced marginal periodontitis.

Different bacterial profiles of subgingival plaque have been described for different clinical categories of marginal periodontitis. In the present investigation the subgingival microflora was studied in young adults with advanced marginal periodontitis. From 12 patients pooled subgingival plaque samples from 5 advanced stages of diseased sites were examined by direct differential phase-contrast microscopy and by cultivation on enriched and selective media. The proportions of the following genera and species were calculated: black-pigmented Bacteroides sp., B. gingivalis, Fusobacterium nucleatum, Veillonella sp., Actinobacillus actinomycetemcomitans, Capnocytophaga sp., Eikenella corrodens, Campylobacter sp., and Candida sp. Spirochete identification was carried out ultrastructurally. Calculated percent minimum similarity levels between the individuals revealed that each patient harbored its characteristic cultivable subgingival microflora different from the other individuals. The spirochetes seemed to constitute the subgingival bacterial group with the smallest variation as certain morphotypes were dominating in almost all individuals.

Adult↗

PubMed1999

Culture of gingival fibroblasts on bioabsorbable regenerative materials in vitro.

BACKGROUND: The use of membranes in guided tissue regeneration (GTR) can limit the apical migration of gingival cells and favor the establishment of new attachment by periodontal ligament fibroblasts. However, gingival recession during healing following GTR has been described as a frequent complication. The purpose of this study was to determine if gingival fibroblasts are affected by the composition of the bioabsorbable membranes used in mucogingival surgery. METHODS: Two type of bioabsorbable regenerative materials were used as cell carriers. Wistar rat gingival fibroblasts (RGF) were obtained from attached gingiva, cut into small fragments, and placed in culture dishes. When confluent, cells were detached using trypsin and identified as "first transferred cells" (P1). At the third passage (P3), cell count, trypan blue exclusion test, acid phosphatase activity, DNA synthesis, phase contrast microscopy, and scanning electron microscopy were performed. The cells were then placed in wells containing the membranes and incubated for 72 hours. RESULTS: When examined under microscopy, the control wells (without membranes) showed one cell type with the elongated appearance characteristic of fibroblasts. The wells with membranes showed an altered cell morphology with a high proportion of cell fragments regardless of the type of membrane used. CONCLUSIONS: These results suggest that cell carrier membranes could affect RGF morphology and thus alter gingival tissue healing following GTR.

Absorbable Implants↗

PubMed2000

The anterior lens capsule used as support material in RPE cell-transplantation.

PURPOSE: To investigate the use of an ocular basement membrane as support material for transplanted porcine RPE cells. METHODS: Porcine RPE cells were grown on bovine corneal extracellular matrix (ECM), isolated bovine- and porcine lens capsules, and tissue culture plastic. Cell density, and cell morphology were studied by phase contrast microscopy and transmission electron microscopy. RESULTS: RPE cells grown on porcine anterior lens capsule and on ECM obtained better morphology and higher final cell density than cells grown on plastic and on bovine anterior lens capsule. It was possible to transplant the porcine anterior lens capsule to the subretinal space in pigs. Within two weeks of observation, the lens capsule was well tolerated in the subretinal space. CONCLUSION: The anterior lens capsule seems to be promising as support material for use in RPE cell-transplantation.

Animals↗