Polarized-light intravital microscopy for study of cochlear microcirculation.
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Animals↗
SEARCH · PubMed Health
Results for “Microscopy, Polarization”
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Pathological alterations in human tendons.
The present authors analyzed the pathological alterations of 1966 tendons examined in the National Institute of Traumatology, Budapest, during the past 18 years. The majority of cases proved to be tendinopathies (hypoxic-degenerative tendinopathy or calcific tendinitis, tendolipomatosis and mucoid degeneration) leading to tendon rupture. The incidence of tendon tumors, foreign bodies, infectious tendon diseases, and other pathological conditions was clearly lower. The methods of tissue preparation and of examination of tendon specimens were also evaluated. Light microscopy was sufficient for the diagnosis of pyogenic tendinitis, tumors, xanthoma, gout, and gangrene. In degenerative tendinopathies and alterations due to hereditary disease, electron microscopy was necessary. Polarization microscopy had a key role in examination of collagen structure and architecture, and identification of foreign bodies in the tendons. Enzyme histochemical and immunohistochemical examination were reliable but not absolutely necessary in the diagnosis of tendon pathology.
Academies and Institutes↗
Polarization and fluorescent microscopy--simple methods for demonstrating pulmonary surfactant lipids.
Birefringence and fluorochrome lipid staining with benzpyrene are demonstrated as simple morphological methods to reveal the presence or absence of surfactant lipids in human newborn lung tissue. Lack of lipid birefringence proves to be an associated finding in the lungs of premature infants with hyaline membrane disease, indicating the possible pathogenetical importance of surfactant deficiency.
Benzopyrenes↗
Preparation and evaluation of diltiazem hydrochloride-Gelucire 43/01 floating granules prepared by melt granulation.
The basic objective of this study was to explore the application of Gelucire 43/01 for the design of multi-unit floating systems of a highly water-soluble drug diltiazem HCl. Diltiazem HCl-Gelucire 43/01 granules were prepared by melt granulation technique. The granules were evaluated for in vitro and in vivo floating ability, surface topography, and in vitro drug release. Aging effect on storage was evaluated using scanning electron microscopy, hot stage polarizing microscopy (HSPM), differential scanning calorimetry (DSC), and in vitro drug release. Granules were retained in stomach at least for 6 hours. Approximately 65% to 80% drug was released over 6 hours with initial fast release from the surface. Surface topography, HSPM, DSC study of the aged samples showed phase transformation of Gelucire. The phase transformation also caused significant increase in drug release. In conclusion, hydrophobic lipid, Gelucire 43/01, can be considered as an effective carrier for design of a multi-unit floating drug delivery system of highly water-soluble drugs such as diltiazem HCl.
Adult↗
[Study of structural changes in muscle fiber contractile proteins using polarization ultraviolet fluorescence microscopy. II. Effect of troponin-tropomyosin complex on the conformation of F-actin in muscle fibers].
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Actins↗
[Panorama replica procedure, a modified method for the description of fine structures in light, polarization and electron microscopy].
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Microscopy↗
Amyloid in adrenal gland pheochromocytomas.
OBJECTIVE: To assess the frequency of amyloid deposits in adrenal gland pheochromocytomas. DESIGN: We evaluated 22 pheochromocytomas using routine hematoxylin-eosin and Congo red stains, polarization microscopy, and electron microscopy. Cases with amyloid were further characterized immunohistochemically. RESULTS: Congo red and polarization microscopy revealed characteristic apple-green birefringence, not abolished by potassium permanganate incubation, in only one neoplasm. In a subset of the remaining tumors, yellow or yellow-green birefringence not characteristic of amyloid was found. We performed electron microscopy on the tumor with characteristic apple-green birefringence and six other neoplasms with abundant yellow or yellow-green birefringence. Amyloid was identified in two tumors, including the Congo red-positive tumor, while the remaining five neoplasms contained collagen. Immunohistochemically, the amyloid deposits were reactive with component P and immunoglobulin kappa- and lambda-light chains in both tumors. Adsorption studies using human urine with monoclonal immunoglobulin kappa- or lambda-light chains abolished reactivity, consistent with specific immunostaining for both light chains. Transthyretin was present in one tumor. The neoplastic cells expressed chromogranin, neuron-specific enolase, and [Leu5]-enkephalin, although the amyloid was not reactive. CONCLUSIONS: We conclude that amyloid is infrequent in pheochromocytomas. Electron microscopic examination is more sensitive than Congo red stain and, in our opinion, is the best technique for the demonstration of amyloid deposits.
Adrenal Gland Neoplasms↗
Sensitivity and specificity of Congo red staining according to Romhányi. Comparison with Puchtler's or Bennhold's methods.
The sensitivity and specificity of various Congo red staining methods is very important in the diagnosis of amyloidosis. When using a less sensitive staining method, some true positive cases of amyloidosis remain undetected. A more highly specific method potentially detects more cases and reveals amyloidosis in an earlier stage of deposition. In this paper, the Congo red staining method according to Romhányi is discussed in comparison with Puchtler's and Bennhold's methods. Using Romhányi's technique, there is no alcoholic differentiation, and thus no dye molecules are washed off the amyloid filaments. The binding of the oriented dye molecules is optimal for polarization microscopy. With this method, the polar hydrophilic mounting medium, gum Arabic is used. Mounted in this carbohydrate-containing, hydrophilic medium, the Congo red molecules are oriented parallel to the surface of the amyloid filaments and the sign is linear positive, corresponding to an additive character of topo-optical staining reactions. Otherwise, the Congo red molecules are oriented perpendicular to the surface of collagen, reducing the intensity of birefringence and even inducing an inversion of the original sign of the collagen birefringence. With alcoholic differentiation, Congo red dye molecules are extracted and this decreases the birefringence of amyloid deposits, i.e. minimal amyloid deposits may be missed. Using the apolar hydrophobic mounting medium, Canada balsam, an axis-parallel arrangement of Congo red dye molecules on the surface of collagen fibers and amyloid will occur, resulting in an additive topo-optical reaction with a green polarization color and a false positive diagnosis of amyloidosis ("phantom amyloidosis").
Amyloid↗
Studies on structural changes of F-actin and myosin in living, intact and damaged muscle fibres by means of polarized ultraviolet fluorescence microscopy.
By means of polarized ultraviolet fluorescence microscopy the structural changes of F-actin and myosin were discovered at the changing a functional state of a living muscle fibre and during spreading degeneration (Zenker's necrosis). The character of conformational changes of F-actin and myosin at activation, contraction, contracture and rigor is similar, but the number of changed macromolecules depends on a fibre state. At fibre local damage in its morphological intact parts there was found an alternation of zones, reflecting two states unusual for a fibre. During spreading degeneration these states transform into irreversible contracture and then into rigor. Similar changes were observed in muscle fibres obtained from denervated muscles.
Actins↗
Imaging of anisotropic cellulose suspensions using environmental scanning electron microscopy.
The effect of concentration on anisotropic phase behavior of acid-hydrolyzed cellulose suspensions has been examined using conventional polarizing microscopy and the novel technique of environmental scanning electron microscopy (ESEM). Microcrystalline cellulose dispersed in water formed biphasic suspensions in a narrow concentration range, 4-12 wt % for a suspension pH of 4, where the upper and lower phases were isotropic and anisotropic (chiral nematic), respectively. It is known from previous work that within the biphasic regime total suspension concentration affects only the volume fractions of the two phases, not phase concentration or interfacial packing. As the total suspension concentration surpassed the upper critical limit (c), however, a single anisotropic phase of increasing concentration was observed. It was evident from polarizing microscopy that the chiral nematic pitch of the anisotropic phase decreased with increasing concentration, which has been attributed to a reduction in the electrostatic double layer thickness of the individual rods, thus increasing intermolecular interactions. Chiral nematic textures were also visible using ESEM. This technique has the advantage of studying individual rod orientation within the liquid crystalline phase as it permits the high resolution of electron microscopy to be applied to hydrated samples in their natural state. To our knowledge this is the first time such lyotropic systems have been observed using electron microscopy.
Anisotropy↗
Correlation of collagen organization with polarization sensitive imaging of in vitro cartilage: implications for osteoarthritis.
OBJECTIVE: Optical coherence tomography (OCT), a new method of high resolution imaging, has shown feasibility for assessing articular cartilage to identify early changes in osteoarthritis (OA) and monitor therapy. OCT is analogous to ultrasound, measuring the intensity of backreflected infrared light rather than sound. The resolution of this technology is up to 25 times higher than existing methods. We investigated the correlation between changes observed by OCT and the degree of collagen organization in OA cartilage. METHODS: Polarization sensitive OCT (PSOCT) imaging was used to assess changes in cartilage collagen organization in vitro. RESULTS: The presence (or absence) of PSOCT changes correlated with collagen organization (or disorganization) on histology as assessed by picrosirius polarization microscopy (no significant difference). In multiple cases, cartilage was abnormal by both PSOCT and polarization microscopy, but was grossly normal by routine staining, showing cartilage thickness > 2 mm and no fibrillations. CONCLUSION: This in vitro study suggests PSOCT changes in cartilage are due to the state of collagen organization. The combination of high resolution structural imaging and birefringence detection make OCT a potentially powerful technology for early assessment of OA.
Cartilage, Articular↗
Interpretation of synovial fluid data.
The routine battery of tests for synovial fluid analysis includes culture and Gram staining, polarizing microscopy, and total WBC and differential counts. If the volume of fluid collected is low, culture and polarizing microscopy have highest priority. Synovial fluid data are diagnostic in only two diseases: septic arthritis and crystal-induced arthritis. In traumatic arthritis, degenerative joint disease, rheumatoid arthritis, and systemic lupus erythematosus, synovial fluid data may provide evidence supporting the diagnosis.
Arthritis↗
Biliary sludge is formed by modification of hepatic bile by the gallbladder mucosa.
BACKGROUND & AIMS: We studied 22 patients with symptomatic microlithiasis to determine whether a contributory role of the gallbladder in the early stage of cholesterol gallstone formation exists. We compared the merits of different methods (ultrasonography and microscopy) and sources (hepatic or gallbladder) of bile samples for diagnosing microlithiasis. METHODS: Paired hepatic and gallbladder bile samples were studied with polarizing microscopy. Nucleation time, bile salts, phospholipid, cholesterol, cholesterol saturation index (CSI), bilirubin, total protein, albumin and mucin concentration were measured. All patients had abdominal ultrasound examination. RESULTS: With polarizing microscopy as the standard, ultrasonography was positive in 13 patients (59%) and negative in 9 (41%). All gallbladder bile samples were positive for microlithiasis by microscopy. Only one hepatic bile sample was positive (P < .0001). There was a disproportional enrichment of total protein, albumin, and mucin (P < .05) in the gallbladder bile and a conversion of bilirubin diglucuronide to monoglucuronide (P < .01). Gallbladder samples had lower CSI but a faster nucleation time (P < .001), which correlates inversely with CSI, total protein, and mucin concentration. CONCLUSION: Biochemical composition and physical chemical behavior of hepatic bile are modified during residence in the gallbladder, contributing to sludge formation. Gallbladder bile has a lower calculated CSI, higher deconjugation of bilirubin, protein and mucin concentration and crystals were present. Hepatic bile samples are inappropriate for microscopic detection of microlithiasis.
Adult↗
Morphological and structural characteristics of diazo dyes at the air-water interface: in situ Brewster angle microscopy and polarized UV/vis analysis.
A morphological analysis is presented for Langmuir films of the diazo dyes Sudan 4 (S4), Sudan 3 (S3), and Sudan red (SR), using Brewster angle microscopy. Stable nonmonomolecular structures are formed at the air-water interface denoted as a plateau in the pressure-area isotherms. Monolayer domains are evident by the contrastless image even before the pressure onset, which grow in size until it reached a condensed monolayer. This behavior resembles that of Langmuir films from simple aromatic fatty acids. Films from all the azo dyes display similar features, according to the surface potential isotherms and in situ polarized UV/vis spectroscopy except for the larger area per molecule occupied by S4 and SR. This is attributed to the presence of CH(3) groups that cause steric hindrance modifying the organization of diazo dye molecules at the air-water interface. UV/vis polarized absorption spectroscopy showed preferential orientation of S4 and S3 on the water surface, while SR molecules lie isotropically. For these three diazo dyes, film absorption was negligible at very large areas per molecule, becoming nonzero only at a critical area coinciding with the onset of surface potential. The critical area is ascribed to the formation of a H-bonded network between water molecules and diazo dye headgroups.
Journal Article↗
Supramolecular behavior of the amphiphilic drug (2R)-2-ethylchromane-2-carboxylic acid arginine salt (a novel PPARalpha/gamma dual agonist).
PURPOSE: This study was conducted to evaluate the aggregation properties of an amphiphilic drug. METHODS: Aggregation of the drug was studied by various methods including phase-contrast and polarized microscopy, spectrophotometry, surface tensiometry, atomic force microscopy, and dynamic light scattering. Lymph-cannulated rats were used to assess fractions of drug that were absorbed into lymphatics. RESULTS: During the pharmaceutical development of an alpha/gamma dual PPAR agonist, a derivative of a chromane-2-carboxylic acid (compound 1), it was discovered that the compound was able to form various aggregates in aqueous media from pH 6.5 to 7.1, whereas aggregating predominantly into micelles at higher pH values. Critical micelle concentrations seemed to be quite low, about 0.25 mM (0.17 mg/mL) in deionized water as determined by spectrophotometric (dye) and surface tensiometry (du Nuoy) methods. Aggregation of compound 1 into large supramolecular aggregates was visualized via phase-contrast microscopy and atomic force microscopy. The observed aggregates ranged from 250 nm to greater than 10 microm in size. Formation of liquid crystalline phases was observed by polarized microscopy as the material was gradually hydrated with water. Lymph studies in rats indicated that up to 6.9% of the orally administered dose of compound 1 in pH 6.5 buffer appeared in lymph, suggesting that supramolecular aggregation may also occur in vivo leading to partitioning between the portal and the lymph routes. CONCLUSIONS: The aforementioned supramolecular aggregation was found to have a profound effect on the pharmaceutical development of the drug and potentially on in vivo absorption of the drug.
Animals↗
Analytical performances of two liquid crystals and their mixture as stationary phases in capillary gas chromatography.
Comparative gas chromatographic applications of two new liquid crystals called LCa and LCb and their equimolar mixture LC(a+b) were investigated. The thermal properties of LCa, LCb and LC(a+b) were established with differential scanning calorimetry (DSC) and polarizing microscopy. Differential scanning calorimetry of LC(a+b) showed that the melting or clearing temperature was intermediate between the corresponding temperatures of the pure compounds. Polarizing microscopy showed that the liquid crystal phase of A + B was nematic. The chromatographic separation abilities LCa, LCb and LC(a+b) were studied using fused silica capillary columns. Interesting analytical performances were obtained: isomeric separation of aromatics, polyaromatics, phenols.
Calorimetry, Differential Scanning↗
Probing the cathepsin D using a BODIPY FL-pepstatin A: applications in fluorescence polarization and microscopy.
Redistribution of cathepsin D, a major lysosomal aspartic endopeptidase, has been related to various pathological progressions during tumor formation and oxidation stress. We have synthesized a fluorescent probe for cathepsin D, where the pepstatin A was covalently conjugated with the BODIPY (Boron dipyrromethene difluoride) fluorophore. In vitro, BODIPY FL-pepstatin A inhibits cathepsin D activity with an IC50 of 10 nM. The nature of its binding to cathepsin D was further characterized using a fluorescence polarization measurement. Results showed that BODIPY FL-pepstatin A selectively binds to cathepsin D at pH 4.5. In fixed cells, BODIPY FL-pepstatin A stained lysosomes, where it co-localized with cathepsin D. This staining was depleted when cells were co-incubated with unlabeled pepstatin A in acidic buffer. In live cells, BODIPY FL-pepstatin A is internalized and transported to lysosomes. The staining in the lysosomes can be competed with unlabeled pepstatin A. These properties, along with the good photostability of the BODIPY FL fluorophore, make this probe a novel tool for the study of the secretion and trafficking of cathepsin D.
Animals↗
Electron microscopy of polar insertions in the lac operon of Escherichia coli.
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Chromosome Mapping↗