[CONTINUOUS VARIATION OF BRIGHT FIELD TO PHASE CONTRAST AND BRIGHT FIELD TO DARK FIELD WITH MAINTENANCE OF EXACT KOEHLER ILLUMINATION].
Explore the source record for details and available documents.
Light↗
SEARCH · Search PubMed
Results for “MICROSCOPY, PHASE CONTRAST”
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Three-dimensional organization of biological tissues: a new method for investigation using scanning electron microscopy.
A new method is described which enables the visualization of hidden cellular surfaces of biological tissues. The method does not depend on the kind of fixative solution, the duration of the fixation, or the quantity of extracellular matrix. The samples are chemically dissociated with potassium ethoxide until the first individual cells appear in the solution. This initial process is followed by phase contrast microscopy, and remaining tissue is investigated with scanning electron microscopy. The method has proved more effective than conventional enzymatic techniques and is a simple, quick, and effective way to investigate the three-dimensional organization of different normal and pathological tissues.
Aorta↗
An immunocytochemical and ultrastructural study of heterogeneity in the human breast carcinoma cell line PMC42.
A recently established human breast carcinoma cell line has been reported to exhibit a number of morphological cell types in monolayer cultures as defined by phase-contrast microscopy. The cell line also produces cords of viable cells floating within the culture medium. Cultures of this cell line, grown in monolayer, on collagen gels, and as floating cords of cells, were studied by transmission electron microscopy and immunocytochemistry. A detailed analysis of the staining pattern obtained with a series of monoclonal antibodies with well-defined human breast epithelial cell specificities and a polyclonal antikeratin antibody showed PMC42 to resemble cultures of both normal human breast and other human breast carcinoma cell lines. The pleomorphic nature of PMC42 cultures was confirmed at an ultrastructural level, and of the eight cell types observed by phase-contrast appearance, seven ultrastructural counterparts were observed. In addition, the presence of intracytoplasmic lumina and overt epithelial differentiation confirm a breast epithelial origin for this cell line.
Breast Neoplasms↗
Dynamic changes in traction forces with DC electric field in osteoblast-like cells.
Primary bovine osteoblasts and human osteosarcoma cells exposed to direct-current electric fields undergo processes of retraction and elongation ultimately resulting in the realignment of the long cellular axis perpendicular to the electric field. The time taken for this reorientation was inversely correlated to field strength within a certain range. Cellular force output during reorientation was analyzed using a simple modification of traction force microscopy. The first detectable reaction was an increase in average traction force magnitude occurring between 10 and 30 seconds of electric field exposure. In the following 2 to 15 minutes traction forces at margins tangential to the electric field decreased below their initial values. Phase-contrast microscopy revealed elongating protrusions at these margins several minutes later. We could not correlate the initial traction changes with any change in intracellular free calcium levels measured using the fluorescent dye Fura-2 AM.
Acrylic Resins↗
Urinary erythrocyte morphology as a diagnostic aid in haematuria.
The value of urinary erythrocyte morphology in diagnosing glomerular and nonglomerular haematuria was studied using phase contrast microscopy in 105 patients with significant haematuria. Fifty-eight (93.6%) out of 62 patients with glomerulonephritis had dysmorphic erythrocytes and 40 (93.1%) out of the 43 patients with nonglomerular disease had isomorphic erythrocytes in the urine. A mixed picture of glomerular and nonglomerular haematuria was seen in 5 patients. The sensitivity was 93.6%, the specificity was 97.7% and the positive predictive value was 98.3% for glomerulonephritis in patients with dysmorphic haematuria. The positive predictive value for a nonglomerular source of bleeding was 96.7% with isomorphic haematuria. It is concluded that phase contrast microscopic examination of erythrocytes in urine is a simple, inexpensive and noninvasive technique that reliably distinguishes between glomerular and nonglomerular bleeding in patients.
Erythrocyte Count↗
Digital image analysis of growth and starvation responses of a surface-colonizing Acinetobacter sp.
Surface growth of an Acinetobacter sp. cultivated under several nutrient regimens was examined by using continuous-flow slide culture, phase-contrast microscopy, scanning confocal laser microscopy, and computer image analysis. Irrigation of attached coccoid stationary-phase Acinetobacter sp. cells with high-nutrient medium resulted in a transition from coccoid to bacillar morphology. Digital image analysis revealed that this transition was biphasic. During phase I, both the length and the width of cells increased. In contrast, cell width remained constant during phase II, while both cell length and cell area increased at a rate greater than in phase I. Cells were capable of growth and division without morphological transition when irrigated with a low-nutrient medium. Rod-shaped cells reverted to cocci by reduction-division when irrigated with starvation medium. This resulted in conservation of cell area (biomass) with an increase in cell number. In addition, the changes in cell morphology were accompanied by changes in the stability of cell attachment. During phase I, coccoid cells remained firmly attached. Following transition in high-nutrient medium, bacillar cells displayed detachment, transient attachment, and drifting behaviors, resulting in a spreading colonization pattern. In contrast, cells irrigated with a low-nutrient medium remained firmly attached to the surface and eventually formed tightly packed microcolonies. It is hypothesized that the coccoid and bacillar Acinetobacter sp. morphotypes and associated behavior represent specialized physiological adaptations for attachment and colonization in low-nutrient systems (coccoid morphotype) or dispersion under high-nutrient conditions (bacillar morphotype).
Acinetobacter↗
Interpretation of airborne asbestos measurements.
Transmission electron microscopy (TEM) is the preferred method of measuring airborne asbestos in buildings, but TEM measurements cannot be used directly in the existing equations relating risk to exposure because the equations are based on measurements made with a different technique--phase contrast microscopy (PCM). Comparison between measurements made by different methods is not simple because the methods differ in the size of particles they can detect, and the relationship between exposure and disease is thought to depend on, among other things, asbestos fiber size. Previous suggestions for converting TEM measurements to PCM equivalents lack generality because they fail to take into account the size distribution of the asbestos particles and the expectation that fiber-size distributions in current nonoccupational environments could differ from the workplaces of the past on which the risk equations are based. A mathematical model is presented for investigating the conversion of airborne asbestos measurements made by one method to an equivalent measurement made by another method. "Equivalent" means having the same potential to cause disease. The model clarifies the issues of concern and suggests approaches for obtaining meaningful conversion factors that will allow TEM measurements to be used in PCM-based risk equations.
Air Pollutants↗
Orientational order of the lamellipodial actin network as demonstrated in living motile cells.
Lamellipodia of crawling cells represent both the motor for cell advance and the primary building site for the actin cytoskeleton. The organization of actin in the lamellipodium reflects actin dynamics and is of critical importance for the mechanism of cell motility. In previous structural studies, the lamellipodial actin network was analyzed primarily by electron microscopy (EM). An understanding of lamellipodial organization would benefit significantly if the EM data were complemented and put into a kinetic context by establishing correspondence with structural features observable at the light microscopic level in living cells. Here, we use an enhanced phase contrast microscopy technique to visualize an apparent long-range diagonal actin meshwork in the advancing lamellipodia of living cells. Visualization of this meshwork permitted a correlative light and electron microscopic approach that validated the underlying organization of lamellipodia. The linear features in the light microscopic meshwork corresponded to regions of greater actin filament density. Orientation of features was analyzed quantitatively and compared with the orientation of actin filaments at the EM level. We infer that the light microscopic meshwork reflects the orientational order of actin filaments which, in turn, is related to their branching angle.
Actins↗
Recovery of neuromuscular junction morphology following 16 days of spaceflight.
It has previously been established that spaceflight elicits alterations in the morphology of the neuromuscular system that includes expansion of the neuromuscular junction (NMJ) and myofiber atrophy. The purpose of this study was to determine the capacity of the neuromuscular system to recover from spaceflight-induced modifications upon return to normal gravity. Soleus muscles were obtained from rats participating in the 16-day Neurolab space shuttle mission at 1 day and 14 days after returning to Earth: solei were also taken at the same time points from ground-based control rats. Cytofluorescent techniques, coupled with confocal microscopy, were used to assess NMJ morphology. Histochemistry, in conjunction with phase contrast microscopy, was employed to examine myofiber size and type. Results indicate that 1 day after landing both pre- and postsynaptic stained areas of the NMJ were significantly (P < or = 0.05) larger in the spaceflight group than in controls. Moreover, significant myofiber atrophy was demonstrated in animals subjected to 0 gravity. By 14 days following return to the Earth, however, NMJ stained areas and muscle fiber size were no longer different from control values at that same interval. These results suggest that the neuromuscular system possesses a robust capacity to recover from spaceflight-induced perturbations upon return to normal gravitational influences.
Adaptation, Physiological↗
Mineral fiber content of lungs in patients with mesothelioma seeking compensation in Québec.
Asbestos fibers (AF) and ferruginous bodies (FB) in lung parenchyma from 50 workers seeking compensation from the Workers' Compensation Board of Québec for pleural or peritoneal mesothelioma were analyzed using transmission electron microscopy (TEM) equipped with energy-dispersive spectrometer (EDS) and phase-contrast microscopy (PCM). These workers had been occupationally exposed in mining and milling activities (12 were from Asbestos Township and 11 from Thetford Mines) and 27 were from other types of industry (asbestos factory, shipyard, etc.). For comparison, analyses of lung tissue at autopsy were done in a group of 49 subjects from a reference population. A 95% confidence interval upper limit of 540 AF < 5 microns/mg and a 95% confidence interval upper limit of 161 AF > or = 5 microns/mg dried lung tissue were found for the reference population. Similarly, a concentration of FB of 142 FB/g constituted the upper limit of detectable FB in the lungs of the reference population. Forty-eight of the 50 workers with mesothelioma had either a ferruginous body or total asbestos fiber count greater than the 95% confidence interval for the reference population; the remaining two had amosite and/or crocidolite concentrations greater than the 95% confidence interval for the reference population. The fiber types were different in the three groups, with the lungs of workers from Thetford Mines containing only chrysotile and tremolite, those from Asbestos Township containing chrysotile, tremolite, amosite, and crocidolite, and those in other industries containing largely amosite and crocidolite. We conclude that in this population of workers seeking compensation for mesothelioma, fiber analysis confirmed occupational asbestos exposure in every case. The fiber types responsible for the tumors are probably different in the three different groups.
Aged↗
A rationale for the management of periodontal diseases: effects of tetracycline on subgingival bacteria.
Microbiologic criteria obtained with phase-contrast microscopy were used in a short-term, double-blind study to measure the effects of systemic tetracycline HCl on subgingival bacterial populations in advanced periodontal pockets refractory to local therapy (repeated scaling, root planing, and the subgingival administration of chemotherapeutic agents [H2O2, NaHCO3, NaCl, MgSO4]). Twenty-one subjects, selected for study, had at least one of the following conditions present after local therapy: spirochetes, motile rods, or crevicular leukocytes greater than or equal to 125 per phase-contrast microscopic field. Tetracycline HCl (1 gm/day for 14 days) was randomly distributed to 11 subjects and a placebo to ten subjects, so that neither the subjects nor investigators were aware of the prescription contents. Evaluations after two weeks disclosed that tetracycline HCl significantly reduced elevated levels of spirochetes, motile rods, and crevicular leukocytes to low or undetectable levels, whereas levels in the placebo subjects remained generally unchanged. The results clearly demonstrate the value of tetracycline HCl as an adjunct to periodontal therapy in reducing remaining suspected periodontopathic bacterial populations in advanced lesions after local therapy of scaling, root planing, and topically applied chemotherapy.
Adult↗
Exposure of UK industrial plumbers to asbestos, Part II: Awareness and responses of plumbers to working with asbestos during a survey in parallel with personal sampling.
Throughout the European Union, millions tonnes of asbestos were used in the manufacture of products for building and for industrial installations. Today, in the UK, it is estimated that over half a million non-domestic premises alone have asbestos-containing materials in them and it is recognized that those working in building maintenance trades continue to be at significant risk. In part II, the awareness of UK plumbers to when they are working with asbestos was investigated and compared with the monitored levels reported in part I. The plumbers were issued by post with passive samplers, activity logs to monitor a working week and a questionnaire. The activity logs were used to assess whether maintenance workers were knowingly or unknowingly exposed to airborne asbestos fibres during a course of a working week. The questionnaire was designed to gather information on their: age, employment status, current and past perception of the frequency which they work with asbestos and knowledge of the precautions that should be taken to limit exposure and risk. Approximately 20% of workers reported on the sample log that they had worked with asbestos. There was a high correlation (93%) between the sampling log replies that they were knowingly working with asbestos and measured asbestos on the passive sampler. However, some 60% of the samples had >5 microm long asbestos structures found by transmission electron microscopy (TEM) analysis suggesting that the plumbers were aware of about only one-third of their contacts with asbestos materials throughout the week. This increased to just over one half of the plumbers being aware of their contact based on the results for phase contrast microscopy (PCM) countable asbestos fibres. The results from the questionnaire found that over half of the plumbers replying thought that they disturb asbestos only once a year and 90% of them thought they would work with asbestos for<10 h year-1. Their expectations and awareness of work with asbestos were therefore far lower than found during the period of monitoring. The response to an open question on the precautions to be taken, found that 61% of plumbers would avoid disturbance and 59% would use respiratory protection if they did disturb asbestos. However, their awareness of the methods and the need to reduce their risk by control of emissions at source was very low and suggests that further awareness raising and training is needed. The survey was carried out prior to the introduction of the duty to manage and gives a useful baseline to assess the impact of regulatory initiatives.
Adult↗
Parameter-optimized digital holographic microscope for high-resolution living-cell analysis.
A parameter-optimized off-axis setup for digital holographic microscopy is presented for simultaneous, high-resolution, full-field quantitative amplitude and quantitative phase-contrast microscopy and the detection of changes in optical path length in transparent objects, such as undyed living cells. Numerical reconstruction with the described nondiffractive reconstruction method, which suppresses the zero order and the twin image, requires a mathematical model of the phase-difference distribution between the object wave and the reference wave in the hologram plane. Therefore an automated algorithm is explained that determines the parameters of the mathematical model by carrying out the discrete Fresnel transform. Furthermore the relationship between the axial position of the object and the reconstruction distance, which is required for optimization of the lateral resolution of the holographic images, is derived. The lateral and the axial resolutions of the system are discussed and quantified by application to technical objects and to living cells.
Algorithms↗
Clonal analysis of expression of epithelial antigens in cultures of normal human breast.
Cells from normal human breast epithelium were cloned in monolayer culture and the clones were stained with monoclonal antibodies. Tissue was from reduction mammoplasty operations. Cloning efficiencies were 5-30%. Two types of clone were identified: 10 to 30% were of relatively spread cells whose boundaries were often difficult to see by phase-contrast microscopy but where they were visible they appeared as dark lines. The edges of the clones usually appeared to be under tension. These clones were stained by two monoclonal antibodies, LICR-LON-M8 and M24, that stain luminal epithelial cells in the intact tissue, but not myoepithelial or stromal cells. Within a clone the cells showed a full range of antigenic phenotypes. This was confirmed for clones grown from single cells that had been isolated manually. The second type of clone was more compact with little evidence of tension at the edges, and cell boundaries were clearly visible and bright under phase contrast. These clones were not stained by antibodies M8 or M24. Both types of clone stained with a third monoclonal antibody that is specific for luminal epithelial cells in the intact tissue, LICR-LON-M18, but the distribution of staining was different in the different types of clone. The simplest interpretation of the two types of clone is that luminal epithelial cells give rise to the spread type of clone while the myoepithelial cells give rise to the more abundant and vigorous compact clones. Alternatively, the compact clones may be from luminal epithelial cells that have lost differentiated characteristics.
Antibodies, Monoclonal↗
Cytotoxic effects of FK506 on human renal proximal tubule cells in culture.
FK506 has been used as the primary immunosuppressive agent administered after a variety of organ transplants, with less reported nephrotoxicity than that of cyclosporine. This study examined in vitro cytotoxicity of FK506 on normal human renal proximal tubule cells. Cytotoxicity was assessed by neutral red inclusion and trypan blue exclusion; morphology was assessed by light and transmission electron microscopy. Neutral red inclusion decreased to less than 10% of the control after 3 days exposure to 200 micrograms/ml FK506. Forty microgram per milliliter FK506 caused a decrease in neutral red inclusion to 61% of the control on Day 7, with recovery to 86% on Day 12. Similarly, trypan blue exclusion decreased to 66% of the control following 7 days exposure to 40 micrograms/ml FK506, and confluency of the monolayer was reduced to 50% as evidenced by phase contrast microscopy. After a 12-day exposure, treated monolayers became more confluent. On ultrastructural examination, FK506-treated cells exhibited increased cytoplasmic vacuolation and lipid inclusion. These data suggest that FK506 is reversibly and mildly toxic to monolayers of human renal proximal tubule cells and are consistent with clinical reports of reversible nephrotoxicity.
Cell Survival↗
Glycerol derivatives of cutin and suberin monomers: synthesis and self-assembly.
Glycerol derivatives of cutin and suberin monomers were synthesized by acid catalysis. Their dispersion in an aqueous solution was examined by phase contrast microscopy, neutron scattering, and solid state NMR. It is shown that the phase behavior strongly depends on the nature of the derivatives forming either lumps of aggregated membranes or well dispersed membranes.
Glycerol↗
Exposure of UK industrial plumbers to asbestos, Part I: Monitoring of exposure using personal passive samplers.
Epidemiological data suggest that there has been and may continue to be a significant risk to maintenance workers, who through their work may disturb asbestos-containing materials (ACM). The sampling and assessment of maintenance workers' exposure is a particular problem because they may not know that they are working with ACM. A strategy to monitor their true exposure has been developed and applied to one group of workers. The asbestos exposure of industrial plumbers was measured using personal passive samplers developed at the Health and Safety Laboratory (HSL). The light-weight samplers, which collect particles by electrostatic attraction, are simple to use and do not require prior knowledge that asbestos is to be disturbed as does conventional sampling. The samplers were issued by post and analysed, after return, using transmission electron microscopy (TEM). The strategy was found to be a reasonably efficient and cost-effective way to obtain data on maintenance worker's exposure to asbestos. The results of the TEM analysis of the passive samplers showed that the percentage of workers exposed to >5 microm long asbestos fibres was 62% in Round 1 and 58% in Round 2. For phase contrast microscopy equivalent (PCME) asbestos fibres, the values were 46 and 29%, respectively. The three samples with the highest numbers of fibres were followed up and were associated with plumbers working in areas which had supposedly been stripped of asbestos just prior to their starting work, suggesting that poor removal, clean-up and clearance practice presents a significant part of the risk to plumbers. Although flow rates will vary with conditions and time, an approximate average sampling rate from previous comparisons was used to calculate the concentration. This gave an average exposure to regulated PCME fibres of 0.009 f ml-1 for amphibole asbestos and 0.049 f ml-1 for chrysotile. The calculate risk based on the PCME fibre types collected and their estimated concentrations, showed that the risk from airborne amphibole fibres was approximately 6 times greater than from chrysotile fibres. If representative, the estimated lifetime risk of death from an asbestos related cancer for an exposure from age 20 for 40 years would be 68 per 100,000, which equates to an annual risk of death of the order of 10 per million.
Air Pollutants, Occupational↗
[Unstained chromosomes. A simple method for observation].
A technique is described by which unstained metaphases may be observed using phase contrast microscopy. The cells are fixed on a cover slip which then is turned over and onto a slide, leaving a thin air layer sandwiched between. Observation in phase contrast shows metaphases comparable to those observed after Giemsa staining.
Chromosomes↗