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PubMed1989

[Application of polarized microscopy and analytic electron microscopy in pneumoconiotic pathologic examination].

38 cases of pneumoconiotic pathologic slides have been reexamined by polarized microscopy, among this, 3 cases (every 1 case of silicosis, asbestosis, cement dusts pneumoconiosis) have investigated by analytic electron microscopy. Authors presented, crystal silica and asbestos fiber may be showing under polarized light, and often associated with carbon dusts or fibrotic proliferation. So that, polarized microscopy may be as a preliminary detect method for silica and uncoated asbestos fiber from histologic slide, according to its dimension, color, lightness, morphologic feature and position were differential diagnosis from other positive substance under routine pathologic examination, further demonstration must to be reduced to ashes and re-observation under polarized microscopy. Also we selected 3 cases of silicosis, asbestosis, cement dusts pneumoconiosis specimen prepared to ultramicrotomic slides and examined by H-800 electron microscopy, when discovered some particles (such as crystal, non-crystal or fibrotic dusts) in cytoplasm of macrophages or interstitial space then used EDAX estimated. Its elements distribution were conformity with dusts (silica, asbestos, cement dusts).

Anthracosilicosis↗
PubMed1995

Localization of Hydrogen Peroxide Production in Pisum sativum L. Using Epi-Polarization Microscopy to Follow Cerium Perhydroxide Deposition.

Cerium is becoming an increasingly popular reagent for histochemical localization of oxidases and phosphatases because it combines directly with reaction products to form fine precipitates of electron-dense materials that can be easily detected using transmission electron microscopy or laser confocal scanning microscopy. We used epi-polarization microscopy to detect cerium perhydroxide deposits formed when H2O2 was produced by diamine oxidase in pea (Pisum sativum L.) epicotyls exposed to exogenous putrescine. Diamine oxidase activity was abundant in cortical cell walls but showed little, if any, association with vascular tissues. Maps of cerium deposition generated using scanning electron microscopy/x-ray microanalysis verified these observations. This study demonstrates the use of epi-polarization microscopy to follow cerium deposition, and the ready accessibility of this microscopy technique should facilitate more widespread use of cerium for plant histochemistry and cytochemistry.

Journal Article↗
PubMed1998

Combination of digital image analysis and polarization microscopy: theoretical considerations and experimental data.

The potentialities of polarization microscopy has been greatly increased by using specific stains for selective enhancement of the optical anisotropy of a macromolecular constituent of cells and tissues. Such stainings have proved to be especially useful in exploring the spatial orientation pattern of the extracellular matrix components. The retardation value, which characterizes quantitatively the degree of submicroscopic orientation, can be measured traditionally with a compensator plate. This technique, however, is time-consuming and greatly dependent on visual judgment. Several attempts have been made to combine digital image analysis and polarization microscopy to improve the measuring technique in unstained structures. In this paper, we summarize theoretical considerations and experimental data to show the advantages and limitations of this methodological approach when using stained and birefringent specimens. The technique we are suggesting is the measurement of the light intensity using a 12 bit cCCD camera attached to a polarized light microscope and digital image analysis system. The theoretical basis is given by the Fresnel equation describing the relationship between light intensity and retardation value. According to this, there is a sin2 function between the light intensity and the retardation value. The same relationship of these two parameters was observed in our experiments on the birefringent extracellular matrix around chondrocytes grown in agarose gel and interterritorial and territorial matrix of canine articular cartilage stained with picrosirius red. Our results suggest that the retardation values can be calculated directly from the light intensity values if the retardation value is lower than lambda/2.

Animals↗
PubMed2004

[The dynamic changes of collagen fiber in quartz-induced pulmonary fibrosis in mice with picrosirius red plus polarized microscopy in vivo].

OBJECTIVE: To investigate the use value of picrosirius red staining plus polarized microscopy to observe the dynamic changes of collagen fiber in lung fibrosis in silicotic mice model. METHODS: The experimental mice were divided into control and quartz groups. 0.2 g/kg weight of quartz was injected intratracheally in quartz group. Lung tissues were collected at the 1st, 3rd, 5th, 7th, 14th and 28th day after injection respectively. Lung tissue slides were stained with picrosirius red. With the aid of polarized microscope, image analysis software, the distribution and change of type I and type III collagen could be qualitatively and quantitatively analyzed. Lung tissue hydroxyproline was determined by chloramines T method. RESULTS: In early stage the predominant increment was type III collagen, but in late stage type I was predominant. The contents of both type collagen tended to increase as postexposure time prolonged. The time course of the ratio of type I to type III showed increasing trend, and there was a statistical significance on day 28 (1.49 +/- 0.39 vs 0.59 +/- 0.24, P < 0.05). The total area of collagen was positively correlated with hydroxyproline concentration of lung tissue (r(2) = 0.928 5, P < 0.01). CONCLUSION: Picrosirius red staining combined with polarized microscopy and digital image processing is a useful method to elucidate collagen accumulation, distribution and subtype ratio in silicosis.

Animals↗
PubMed1977

[On applicabilities of polarization microscopy in diagnostic histopathology (author's transl)].

Apart from its importance in ultrastructure research polarization microscopy turns out to be a valuable additional method in the daily hitological diagnostics of pathologist. By means of some examples the applicabilities of the polarization microscopy in this field are demonstrated. After some physical bases and practical hints for the use of polarization microscopes some staining reactions leading to an optical anisotropic effect are reported. Morever, it is reffered to the demonstration and characterization of crystals in biological fluids and tisdues. In our opinion, simple polarization optics should belong to the optical equipment of each pathologist.

Aorta↗
PubMed2006

A new approach for improving the birefringence analysis of dental enamel mineral content using polarizing microscopy.

The main problem in interpreting birefringence of dental enamel under polarizing microscopy is the lack of physical constants able to allow the Wiener equation to be applied directly to the composition of such tissue. The present study introduces a new approach to circumvent this constraint. Because the nonmineral phase of enamel is heterogeneous, its refractive index can be computed in terms of its components (namely, water, which is partially replaced by the immersion medium, and organic matter), thereby providing a more acceptable refractive index to be used in the Wiener equation. Furthermore, the enamel mineral volume is ordinarily calculated on the basis of the density 3.15 g cm(-3). The density 2.99 g cm(-3) has been, however, reported to be more accurate for enamel hydroxyapatite, so enamel mineral volumes from selected published data were converted using such a density. The birefringence of mature enamel computed by the Wiener equation, taking into account the above refinements, matched, for the first time, published experimental birefringence values. The theoretical water and organic contents were also consistent with published experimental data. Thus, a direct application of the Wiener equation to the enamel composition has now been achieved. It is speculated that quantitative data on the mineral, the water and the organic contents of mature dental enamel can be derived from interpretation of birefringence in two immersion media (obtained before and after extraction of the organic matter) with this new approach.

Biometry↗
PubMed1991

Crystalluria determined by polarization microscopy. Technique and results in healthy control subjects and patients with idiopathic recurrent calcium urolithiasis classified in accordance with calciuria.

A retrospective study was done on the nature and degree of crystalluria in spontaneously voided fasting and postprandial urine of patients with recurrent idiopathic calcium urolithiasis (RCU) divided into normocalciuria (20 males, 20 females) and hypercalciuria patients (20 males, 20 females), and controls (20 males, 20 females). The crystals were obtained using a filter technique and identified by microscopy. In addition, individual data, clinical chemistry variables and indices reflecting the risk of calcium phosphate and calcium oxalate crystallization were evaluated. In contrast to findings of other investigators of crystalluria we observed only a few crystals on the filters. The most frequently occurring phases were (in this order) a urate-containing phase (tentatively termed uric), an amorphous calcium phosphate phase (tentatively termed isotropic) and a phase of spheroid-like particles, not yet definitely characterized (tentatively termed spheroid). Calcium oxalate crystals were found only exceptionally. There was no relationship between the degree of calciuria (normo- versus hypercalciuric RCU) and crystalluria. Among RCU, males generally had a predominance of the isotropic, females of the spheroid phase, as compared with controls. Also, RCU females were generally obese, and their spheroid score and lean body mass correlated negatively and significantly. The calcium phosphate and calcium oxalate risk indices were always low in normal individuals, higher in RCU. Patients of both sexes with urinary stones had normal parathyroid gland function, but higher total calcium in fasting serum and higher urinary pH as compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1982

Polarization microscopy and microspectrophotometry of Sirius Red, Picrosirius and Chlorantine Fast Red aggregates and of their complexes with collagen.

A detailed quantitative analysis of the anisotropic properties of Sirius Red F3B, Picrosirius, and Chlorantine Fast Red crystals, and of their complexes with a macromolecularly oriented protein either in a pure form or as part of a tissue structure was carried out. Collagen I was used as the protein model. Linear dichroism and dispersion of birefringence were investigated in dye aggregates, in stained filaments of collagen I and in collagen bundles in sections of tendon. A positive linear dichroism, the characteristics of which varied as a function of the dye type used, was demonstrated for the dye aggregates and stained substrates. However, even thin regions of the stained tendon collagen bundles showed very high absorbances, differing from the pattern reported previously for collagen stained with another sulphonated azo dye, Xylidine Ponceau. Consequently, not all these dyes enable protein concentration and orientation to be determined in collagen-containing structures. From the linear dichroism patterns it is assumed that the long axis of the molecules of these azo dyes is mostly parallel to that of filaments of pure collagen I and statistically parallel to the long axis of collagen bundles of tendon sections. The dye aggregates and, stained pure collagen I and tendon collagen bundles exhibited birefringent images with interference colours that varied as a function of thickness and packing state of the preparations, which is in agreement with reports in the literature. The optical retardations of the collagen bundles increased by a factor of 5-6 times after staining with Picrosirius. From data on form dichroism it is concluded that when studying the macromolecular orientation of collagen preparations stained with azo dyes, the choice of the mounting medium deserves consideration.

Azo Compounds↗
PubMed1989

[Spheno-occipital synchondrosis--a fluorescence and polarization microscopy study in Cercopithecus aethiops monkeys].

On 17 Cercopithecus aethiops monkeys we investigated with the method of the polychromic sequential dye marking system the histomorphology as well as the dynamics of growth and calcification of the spheno-occipital synchondrosis. The age of the animals ranged from change to teeth to late adolescence. The animals had been divided into four different groups: I: late change of teeth; II: young adult; III: fully grown; IV: late adolescence. In the first group the spheno-occipital synchondrosis showed no ossification in the gap which is filled with cartilage. It showed the characteristic structure with a central zone of equally distributed chondrocytes. Adjacent to this we found a zone of proliferation cell hypotrophy and cell degeneration. With increasing age there is decrease of the density of cells (groups II to IV) and after change to teeth the synchondrosis starts to ossify (group II). Due to the ossification the synchondrosis subdivides into different cartilage regions. We found that the closing of the synchondrosis started in the cranial region and progressed toward the caudal region. During this procedure the synchondrosis never ossified completely. Several cartilage regions persisted uncalcified until late adolescence. Interstitial growth of the synchondrosis was found until the end of the change of the teeth (group I). This growth which was always found in a sagittal direction ceased after bony connections had been formed between both poles of the synchondrosis. The sphenoidal and occipital pole of the synchondrosis showed equal growth potential.

Aging↗
PubMed1981

Structure and porosity of human cervical enamel studied by polarizing microscopy and transmission electron microscopy.

Dehydration by alcohols and concentrated solutions of highly soluble salts caused refractive changes in enamel which were enhanced in the cervical regions. Electron microscopy showed crystals in cervical enamel to vary from normal orientation, size and habit to marked disorientation and reduced size. Consequent increases in surface area and inter-crystalline volume could account for the exaggerated optical behaviour. The changes in optical properties of enamel occurred only when the osmotic pressures of the imbibing solutions exceeded 500 atm. It is proposed that the internal pore system may take up water freely but exclude large hydrated molecules and ions which thereby create a negative pressure on the water contained in these pores. Once this negative pressure exceeds the tensile strength of the water-pore system, molecules may be torn from their attachment sites, and possibly from each other, so that a sudden evacuation of the pores is accompanied by a simultaneous reduction in refractive index. It is likely that such sudden evacuation would alter the potential of the pore surfaces to react with small molecules and ions subsequently invading the tissue.

Absorption↗