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PubMed1980

[Nomarski interference microscopy: use of combined contrast uranyl acetate-PTA for in vitro observation of monolayer cultures].

In this paper is reported a technique for the fixation and staining of cellular monolayers in vitro for observation by the interferential microscope according to Nomarski. This method includes the use of glutaraldehyde at 2.5% in phosphate buffer 0.1 M pH 7.5 and the sequential use of Uranil acetate (0.9% in absolute alcohol) and phosphtungstic acid (1% in absolute alcohol). This study has been conducted on 3 cellular lines: PC12 (rat pheocromocytoma), R.P.C. (rat pineal cells), primary culture obtained from a human carcinoma of the uterus.

Acetates↗
PubMedDate not supplied

Comparison of buccal and nasal epithelial cells using a new cell development index and quantitative interference microscopy.

Cell area, cell dry mass, and three summary indices for tissue maturation were obtained for human buccal and nasal cells viewed in smears (five donors). The means for cell area and dry mass in the buccal smears were 3816 X 10(-8) cm2 and 2294 pg (1 pg=1 X 10(-12) g), respectively. The correlation coefficient for means of cell area and dry mass was r=0.03. The means for the three maturation indices used to characterize the cells viewed in buccal smears were: Cell Development Index (CDI), 486; Maturation Index (MI) 50.6; and Karyopyknotic Index (KPI), 0.028. The correlation coefficients for means were: CDI vs. MI, r=0.88; CDI vs. KPI, r=0.71; and MI vs. KPI, r=0.73. In the buccal smears, the cell types found most frequently were intermediate with round or oval nuclei, which altogether averaged about 90% of the total smear composition, while superficial cells (pyknotic ghost, and anucleate) comprised less than four per cent. In nasal smears, the means for cell area and dry mass were 867 X 10(-8) cm2 and 623 pg, respectively. The correlation coefficient for means of cell area and dry mass was r=0.74. The means for the maturation indices were: CDI, 816; MI, 91.2; and KPI, 0.79. The correlation coefficients for means were CDI vs. MI, r=0.93; CDI vs. KPI, r=0.97; and MI vs. KPI, r=0.85. In nasal smears, the anucleate cell type was found most frequently (about 75%) while intermediate cells with round or oval nuclei comprised approximately 13% of the total smear composition. When donor means for cell area and dry mass of buccal cells were combined for study, they were highly correlated (r=0.88). Similarly, when the maturation indices for buccal and nasal cells were combined for study, they were highly correlated: CDI vs. MI, r=0.99; CDI vs. KPI, r=0.99; and MI vs. KPI, r=0.99. Of the three indices, we believe that the CDI is the index best suited for use in studies of cell development because it is based on an inferred sequence of cellular and nuclear changes occurring during the maturation of epithelial cells. It provides a more detailed description of the cells observed in the smears. It also correlates highly in KPI and MI used in clinical and research efforts.

Cell Nucleus↗
PubMed1975

Development with age of human articular cartilage surface structure. A survey by interference microscopy of the lateral femoral condyle.

The weight-bearing surfaces of the lateral femoral condyles taken from twenty normal human cadavers aged 0-47 years have been examined by reflected light interference microscopy (RLIM) and by scanning electron microscope (SEM). The surfaces appeared normal by naked eye examination. The presence of both 200-400 mum diameter secondary undulations and small ovoid 20-45 mum diameter tertiary hollows was confirmed in all specimens using both techniques. Measurements by RLIM showed that the tertiary hollows increase significantly in depth and diameter with increasing age. A further order of quaternary surface irregularities was shown. Small irregular ridges, 130-275 nm deep and 1-4 mum diameter, were found with increasing frequency on specimens obtained from persons aged 21 years. These quaternary irregularities are thought to be due to exposure of superficial fibre bundles after the loss, with age, of surface ground substance.

Adolescent↗
PubMed1969

Spatial patterns of threadlike elements in the axoplasm of the giant nerve fiber of the squid (Loligo pealii L.) as disclosed by differential interference microscopy and by electron microscopy.

The giant nerve fiber of the squid (Loligo pealii L.) has been investigated in situ, and in fresh and fixed preparations, by differential interference microscopy and electron microscopy. A continuous, three-dimensional network, composed of threadlike elements, was disclosed in the axoplasm. The threadlike elements in the axoplasm are twisted as a whole into a steep, right-handed helix. In a peripheral ectoplasmic region, the elements are more parallel to one another and more densely packed than in a central endoplasmic core. The threadlike elements can be resolved into a hierarchy of decreasing order of size. Successive levels of the hierarchy are formed by the association of smaller elements into larger ones. The following levels in the hierarchy of network elements have been distinguished: 1-3-micro-wide threads, 0.1-0.35-micro-wide strands, and 70-250-A-wide unit-filament strands. The differential interference microscope selects, from the network, threads oriented at a specific angle to the long axis of the axon. The specific angle depends upon the orientation of the long axis of the axon relative to the direction of shear. It is postulated that the network configuration is expressed in the solid-state properties of the axoplasm essential for the normal functioning of the nerve fiber.

Animals↗
PubMed1978

The surface structure of ageing human articular cartilage: a study by reflected light interference microscopy (RLIM).

The surface contours of the weight-bearing area of the left lateral femoral condyle have been examined in vitro in 26 cadavers by reflected light interference microscopy over the age range 0--47 years. The presence of 4 orders of surface irregularity was confirmed. In addition, measurement of the depth, diameter and frequency of the tertiary order of surface hollows showed that while none of these variables was sex-associated, age-related changes in the variables were present. Multiple regression analysis showed that these changes could be considered in terms of periods of maturation (0--21 years) and ageing (22--50 years) of the cartilage. Focal changes in the surface were observed with advancing age. These changes appeared to be regressive and possibly represent an early stage in cartilage fibrillation.

Adolescent↗
PubMed1989

Fluid distribution in pork, measured by x-ray diffraction, interference microscopy and centrifugation compared to paleness measured by fiber optics.

Moderately PSE (pale, soft, exudative) and moderately DFD (dark, firm, dry) pork was examined by x-ray diffraction for interfilament separation, by differential interference contrast microscopy for interfiber area, and was centrifuged to measure water holding capacity (WHC). Internal reflectance spectra were measured by fiber optics. For PSE to DFD pork, filament separation ranged from 39 to 48 nm, interfiber area from 42 to 3%, and WHC from 49 to 64%, respectively. The correlation of reflectance with interfilament separation varied considerably with wavelength (reaching r = -.83 at 680 nm, P less than .005). The correlation of reflectance with interfiber area was more uniform across the spectrum (reaching r = .90 at 450 nm, P less than .005), as was the correlation of reflectance with WHC (reaching r = -.80 at 400 nm, P less than .005). At 24 h postmortem, fiber-optic spectrophotometry may be used as a rapid, nondestructive method to predict WHC and potential fluid losses from commercial pork with a moderate range from PSE to DFD. Interfiber area was correlated negatively with filament lattice area and WHC, but no significant correlation was found between filament lattice area and WHC. Filament separation was decreased only slightly by centrifugation. These results indicate that at 24 h postmortem the extra fluid released from PSE pork already has been lost from the myofilament lattice and is awaiting release from compartments downstream such as interfiber and interfascicular spaces.

Animals↗
PubMed1985

Morphology of Giardia agilis: observation by scanning electron microscopy and interference reflexion microscopy.

The flagellated protozoan, Giardia agilis, was isolated from tadpole small intestine and examined by scanning electron microscopy and interference reflexion microscopy. The general morphology of the G. agilis trophozoite is similar to G. muris and G. duodenalis, but with modifications that reflect its elongated form. Interference reflexion microscopic analysis of attachment of G. agilis reveals a pattern of focal contacts by the lateral crest of the ventral disc, the ventrolateral flange, the lateral shield, and by numerous microvillus-like appendages found along the lateral border of the trophozoite. The pattern of focal contacts was observed to be dynamic; trophozoites were observed to make and break the focal contacts in a relatively short time and to glide along the surface of the substratum without breaking focal contacts.

Animals↗
PubMed1988

[Interference microscopy of human dental enamel at various age periods].

Investigation of the human tooth enamel by means of interference contrast makes it possible to reveal certain peculiarities of the prismatic structure in various age groups. For example, in children the enamel has specific porosity of the superficial layers, that is peculiar for teeth with a delayed eruption. With age the enamel becomes more "homogeneous", amount and size of the pores decrease, aprismatic areas in the superficial layer are found more often. Erased teeth are characterized with formation of microdefects, destruction of prisms on the enamel surface and at the same time with increased mineralization of the subsuperficial layer. Thus, age changes of the enamel are of adaptive character--consolidation of structural elements promotes increasing resistivity of the teeth to influence of pathological factors.

Adolescent↗
PubMed1981

Topographic examination of sister chromatid differential staining by Nomarski interference microscopy and scanning electron microscopy.

BrdU-substituted Chinese hamster chromosomes were treated with a hog Na2HPO4 solution and stained with Giemsa to produce sister chromatid differential staining (SCD). The process of SCD was examined with the Nomarski differential interference microscope and the scanning electron microscope. After the Na2HPO4 treatment alone, unifilarly BrdU-substituted (TB) chromatids appeared somewhat more severely collapsed than the bifilarly substituted (BB) chromatids. Subsequent Giemsa staining, however, brought about pronounced piling up of the Giemsa dye on the TB-chromatids but not on the BB-ones, causing highly distinct differential Giemsa staining as well as a marked differentiation in surface topography between the sister chromatids. Removal of the Giemsa dye from the differentially Giemsa stained chromosomes resulted in a disappearance of such a pronounced topographic differentiation.

Animals↗
PubMed1983

The effect of primycin on the intracellular monovalent ion and water contents of rat hepatocytes as revealed by energy dispersive X-ray microanalysis and interference microscopy.

Using energy-dispersive X-ray microanalytic and interference microscopic techniques, the intracellular concentration of the monovalent ions (Na+, K+, Cl+) as well as the intracytoplasmic and intracellular water contents were studied in normal and adrenalectomized rat hepatocytes with and without primycin treatment. Although primycin influenced significantly only the intracellular potassium content of the adrenalectomized group, it exerted a marked influence on the intranuclear water content in both the normal and adrenalectomized rats. The intranuclear water content increased significantly in the primycin-treated animals. The conclusion is drawn that the increased level of hydration of the nuclear substances reflects a 'decondensation' of the chromatin which on the other hand, may represent the basis for the various effects of primycin on the induction of certain hepatic enzymes.

Adrenalectomy↗