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At least 19 recordsLinked to original sources
PubMed1998

The observation of the local ordering characteristics of spermidine-condensed DNA: atomic force microscopy and polarizing microscopy studies.

Condensation of DNA by multivalent cations can provide useful insights into the physical factors governing the folding and packaging of DNA in vivo. In this work, local ordered structures of spermidine-DNA complexes prepared from different DNA concentrations have been examined by using atomic force microscopy (AFM) and polarizing microscopy (PM). Two types (I and II) of DNA condensates, significantly different in sizes, were observed. It was found that for extremely dilute solutions (DNA concentrations around 1 ng/microl or below), the DNA molecules would collapse into toroidal structures with a volume equivalent to a single lambda-DNA (type I). In relatively dilute solutions (DNA concentrations between 1 and 10 ng/microll), a significantly larger structure of multimolecular toroids (circular and elliptical, type II) were formed, which were constructed by many fine particles. Measurements show that the average diameter of these fine particles was similar to the outer diameter of the monomolecular toroids observed in extremely dilute solutions, and the thickness of the multimolecular toroids had a distribution of multi-layers with height increments of 11 nm, indicating that the multimolecular toroidal structures have lamellar characteristics. Moreover, by enriching the DNA-spermidine complexes in very diluted solution, branch-like structures constructed by subunits were observed by using AFM. The analysis of the pellets in polarizing microscopy reveals a liquid-crystal-like pattern. These observations suggest that DNA-spermidine condensation could have multiple stages, which are very sensitive to the DNA and spermidine concentrations.

Bacteriophage lambda↗
PubMed1986

Studies of otoconial development in a "giant-crystal" strain of chicks using scanning electron microscopy, polarized light microscopy, and X-ray crystallography.

Otolith formation was studied in a mutant strain of low-fertility Delaware chicks which exhibit an otolithic defect. In all chicks of this strain, otoliths were present as a fused crystal mass which contained abnormally large (giant) otoconia. Studies of the formation of such otoliths during embryonic development revealed that from the very earliest stages the otoconia were much larger than normal, and in the saccular and utricular otoliths formed a fused mass. These results are interpreted as supporting a hypothesis of the de novo formation of giant otoconia in this giant-crystal strain as opposed to the recrystallization hypothesis proposed for other, dissimilar mutant mammals and birds which also produce giant otoconia.

Animals↗
PubMed1985

Modified procedure for the quantitative estimation of pore volumes in carious dental enamel by polarizing microscopy.

The polarizing microscope was used to make two estimates of pore-volume fraction in carious enamel. Estimates from form birefringence were always much lower than estimates from the intrinsic birefringence of the enamel solid and were too low for carious enamel. The discrepancy cannot be explained except in part as being due to re-precipitation of mineral during lesion formation. The discrepancy may arise largely from the unjustified assumption that the whole tissue acts as a Wiener composite body. Form birefringence is probably produced only by a fraction of the tissue. A model which reconciles the two pore-volume estimates and enables calculation of the composite body fraction and of the volume fractions of solid within this fraction and in the tissue as a whole is described. It appears that polarizing microscopy will require the conjoint application of other techniques for quantitative estimation of pore-volume fractions.

Birefringence↗
PubMed1997

Histologic analysis of thermal effects of laser thermokeratoplasty and corneal ablation using Sirius-red polarization microscopy.

PURPOSE: To evaluate how well several histologic techniques differentiate degrees of thermally induced changes in corneal tissue after laser thermokeratoplasty (LTK) or corneal ablation. SETTING: Medical Laser Center Lübeck, Germany. METHODS: Corneas of freshly enucleated porcine eyes were treated with a continuous wave laser diode (1.86 microns) and a pulsed chromium-thulium-holmium: YAG laser (2.1 microns) to produce LTK lesions or ablated with a Q-switched and a free-running chromium-erbium: YSGG laser (2.70 microns), a free-running erbium: YAG laser (2.94 microns), and an argon-fluoride excimer laser (193 nm). The lesions were evaluated by light microscopy (LM) (hematoxylin and eosin, Azan, van Gieson's, and Masson-Goldner's trichrome stains), transmission electron microscopy (TEM), and polarization microscopy after Sirius-red staining. Sirius-red, a strongly elongated, birefringent molecule binding parallel to collagen molecules, was used to enhance corneal birefringence. RESULTS: With routine LM, it was difficult to discriminate the degrees of thermal alterations in LTK lesions. Combined Sirius-red staining and polarization microscopy distinguished between a strongly coagulated central zone and the transition zone to normal tissue. Sirius-red uptake was increased in both zones, reflecting the availability of new binding sites. The central zone appeared darker under polarization than normal collagen because of a loss of birefringence. Intrinsic birefringence was greatly reduced; however, form birefringence partly remained as long as some collagen fibrils were intact. In the center of very strong lesions, where the collagen was hyalinized, birefringence was completely lost because of the complete disintegration of the fibrillar structure, which was visible under TEM. The transition zone toward normal cornea showed increased birefringence because the natural birefringence was largely preserved and enhanced by the increased Sirius-red uptake. Mechanical stretching between neighboring LTK lesions was manifested by increased birefringence. CONCLUSION: Sirius red offered an improved and simple histologic method for analyzing thermal collagen changes. It may contribute to a better understanding of the working mechanisms of LTK and improve analysis of thermal effects in corneal ablation.

Animals↗
PubMed1988

Sensitive detection of immunogold-silver staining with darkfield and epi-polarization microscopy.

We evaluated the contribution of darkfield and epi-polarization microscopy to the detection of leukocyte cell surface antigens with immunogold-silver staining (IGSS). Lymphocyte cell surface differentiation antigens were labeled with monoclonal antibodies and IGSS as described for brightfield microscopy. In darkfield and epi-polarization microscopy the labeling appeared as bright spots on a dark background. The sensitivity of detection was much higher than that of brightfield microscopy. Sixteenfold higher dilutions of the monoclonal antibody could be used to detect all cells expressing the antigen in the cell suspension. However, non-specific staining was also better visualized. The latter could be reduced to a level comparable to that of brightfield microscopy only by use of weaker labeling conditions. A 25% reduction of the silver enhancement time was necessary for this purpose. However, these weaker labeling conditions also reduced the intensity of the specific staining. Therefore, the efficiency of IGSS, as detected with darkfield and epi-polarization microscopy, was only fourfold greater than that found with brightfield microscopy or that of an immunofluorescence procedure. Especially in combination with transmitted light, to improve cell identification, epi-polarization microscopy is a reliable and sensitive method for detection of immunogold-silver-labeled cell surface antigens for diagnostic and research purposes.

Antibodies, Monoclonal↗
PubMed1988

The effect of speed of deoxygenation on the percentage of aligned hemoglobin in sickle cells. Application of differential polarization microscopy.

We have used differential polarization microscopy, which provides images of linear dichroism, to measure the percentage of aligned hemoglobin (Hb) in 1086 deoxygenated red blood cells from subjects with sickle cell anemia. The percentage was found to be only slightly dependent on the speed of deoxygenation, thus showing that the percentage of aligned Hb was thermodynamically controlled (as has been found previously for the percentage of polymerized Hb in vitro). A slight decrease in the percentage of aligned Hb due to increasing speed of deoxygenation is primarily due to the increase in the number of cells containing no detectable aligned Hb. This class of cells was also the most variable between the different subjects studied. We were able to identify two other groups of cells that contain different numbers of domains of aligned Hb and show that these groups contain statistically different percentages of aligned Hb. The differences between these classes of cells was shown to be primarily due to different numbers of initial nucleation sites within each cell. It appears that the presence of preformed nucleation sites within cells at ambient oxygen tensions results in the thermodynamic control of aligned Hb polymer.

Algorithms↗
PubMed2001

Modulated polarization microscopy: a promising new approach to visualizing cytoskeletal dynamics in living cells.

In an effort to visualize cytoskeletal filaments in living cells, we have developed modulated polarization microscopy. Modulated polarization microscopy visualizes cytoskeletal filaments based on their birefringence but differs from the standard polarization microscopy by exploiting the angle dependence of birefringence. A prototype instrument has been developed using two Faraday rotators under computer control to change the angle of plane polarized light at a known rate. By placing one Faraday rotator before and one after the specimen, rotation produced by the first Faraday rotator is cancelled by the second. This allows the use of fixed polarizer and analyzer in a crossed configuration and continuous imaging of the specimen between crossed polarizers. The variation in polarization angle of light illuminating the specimen causes birefringent elements to oscillate in brightness. Images acquired as polarization angle is varied are then processed by a Fourier filter image-processing algorithm. The Fourier filtering algorithm isolates those signals that vary at the proper rate, whereas static or random signals are removed. Here we show that the modulated polarization microscope can reveal cytoskeletal elements including stress fibers and microtubules in living cells.

Animals↗
PubMed2003

Room temperature measurements of the 3D orientation of single CdSe quantum dots using polarization microscopy.

Simple far-field emission polarization microscopy reveals that the emission transition dipole of CdSe colloidal quantum dots (QDs) is twofold degenerate at room temperature. We measure, model, and compare polarization anisotropy statistics of CdSe QDs and DiI (a one-dimensional emitter). We find excellent agreement between experiment and theory if the transition dipole of CdSe QDs is assumed to be twofold degenerate. This implies that the three-dimensional orientation of the unique crystal axis in QDs can be determined at room temperature with polarization microscopy. We describe an optical setup to measure four polarization angles of multiple single QDs simultaneously and in real time (approximately equal to 16 Hz). We use this setup in a proof-of-concept experiment to demonstrate that the rotational motion of QDs can be monitored in various host matrices.

Journal Article↗
PubMed1978

Fibrillar bodies in leukemic cells revealed by polarization microscopy.

An optical polarizing microscope with a good coefficient of extinction permits the visualization of the cytoplasmic fibrillar body in living preparations and smears of leukemic cells (human leukemias and the L 5222 experimental leukemia). These inclusions are not visible by phase contrast microscopy nor in fixed and stained smears. The detection in living cells of fibrillar bodies makes it possible to study directly the conditions for their formation and their reaction to the effect of certain drugs.

Actin Cytoskeleton↗
PubMed1996

The central odontogenic fibroma and the hyperplastic dental follicle: study with Picrosirius red and polarizing microscopy.

The central odontogenic fibroma (COF) is an uncommon odontogenic tumor in which the microscopic appearance is similar to that of the hyperplastic dental follicle (HDF). Distinction between these lesions is based mainly on their clinical and radiologic features. The collagen of 5 cases of COF and 13 cases of HDF was studied histochemically by staining sections with Picrosirius red and examining them by polarizing microscopy. Polarization colors of the collagen fibers of the lesions were recorded according to their width. While no differences were seen between the polarization colors of thin fibers (<0.8 mm) in both lesions, the polarization colors of thick fibers (1.6-2 mm) of COF were significantly more green and greenish-yellow as compared with those of HDF. These findings suggest that the Picrosirius red polarization method can be used as a diagnostic tool to differentiate between COF and HDF..

Azo Compounds↗