Optical imaging of skin pathology with a polarized light camera

Mature human dental enamel has presented investigators with several difficulties in ultramicrotomy of specimens for electron microscopy due to its high degree of mineralization. This study explores the possibility of combining ion-milling and high voltage electron microscopy as a means of circumventing the problems of ultramicrotomy.A longitudinal section of an extracted human third molar was ground to a thickness of about 30 um and polarized light micrographs were taken. The specimen was attached to a single hole grid and thinned by argon-ion bombardment at 15° incidence while rotating at 15 rpm. The beam current in each of two guns was 50 μA with an accelerating voltage of 4 kV. A 20 nm carbon coating was evaporated onto the specimen to prevent an electron charge from building up during electron microscopy.

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Identification of calcium oxalate crystals in dried or fixed tobacco leaf

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111550 ◽

1984 ◽

Vol 42 ◽

pp. 288-289

Author(s):

Vicki L. Baliga ◽

Mary Ellen Counts

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Calcium Oxalate ◽

Growth And Development ◽

Polarized Light ◽

Calcium Oxalate Crystals ◽

X Ray Diffraction ◽

X Ray ◽

Transmission Electron ◽

Scanning Electron

Calcium is an important element in the growth and development of plants and one form of calcium is calcium oxalate. Calcium oxalate has been found in leaf seed, stem material plant tissue culture, fungi and lichen using one or more of the following methods—polarized light microscopy (PLM), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and x-ray diffraction.Two methods are presented here for qualitatively estimating calcium oxalate in dried or fixed tobacco (Nicotiana) leaf from different stalk positions using PLM. SEM, coupled with energy dispersive x-ray spectrometry (EDS), and powder x-ray diffraction were used to verify that the crystals observed in the dried leaf with PLM were calcium oxalate.

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Differential polarization imaging of sickled cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102390 ◽

1988 ◽

Vol 46 ◽

pp. 62-63

Author(s):

Marcos F. Maestre

Keyword(s):

Optical Properties ◽

Theoretical Approach ◽

Polarized Light ◽

Polarization Microscopy ◽

Spectroscopic Techniques ◽

Polarization Imaging ◽

Circularly Polarized Light ◽

Circularly Polarized ◽

Microscopic Scale ◽

Chiral Structures

Recently we have developed a form of polarization microscopy that forms images using optical properties that have previously been limited to macroscopic samples. This has given us a new window into the distribution of structure on a microscopic scale. We have coined the name differential polarization microscopy to identify the images obtained that are due to certain polarization dependent effects. Differential polarization microscopy has its origins in various spectroscopic techniques that have been used to study longer range structures in solution as well as solids. The differential scattering of circularly polarized light has been shown to be dependent on the long range chiral order, both theoretically and experimentally. The same theoretical approach was used to show that images due to differential scattering of circularly polarized light will give images dependent on chiral structures. With large helices (greater than the wavelength of light) the pitch and radius of the helix could be measured directly from these images.

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High-resolution measurement of birefringent fine structure in living cells using a new polarized light microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100136647 ◽

1995 ◽

Vol 53 ◽

pp. 54-55

Author(s):

Rudolf Oldenbourg

Keyword(s):

Light Microscope ◽

Fluorescent Dyes ◽

Polarized Light ◽

Processing System ◽

Video Camera ◽

Molecular Organization ◽

Computer Assisted ◽

Image Recording ◽

Birefringent Crystal ◽

Technological Advances

The recent renaissance of the light microsope is fueled in part by technological advances in components on the periphery of the microscope, such as the laser as illumination source, electronic image recording (video), computer assisted image analysis and the biochemistry of fluorescent dyes for labeling specimens. After great progress in these peripheral parts, it seems timely to examine the optics itself and ask how progress in the periphery facilitates the use of new optical components and of new optical designs inside the microscope. Some results of this fruitful reflection are presented in this symposium.We have considered the polarized light microscope, and developed a design that replaces the traditional compensator, typically a birefringent crystal plate, with a precision universal compensator made of two liquid crystal variable retarders. A video camera and digital image processing system provide fast measurements of specimen anisotropy (retardance magnitude and azimuth) at ALL POINTS of the image forming the field of view. The images document fine structural and molecular organization within a thin optical section of the specimen.

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Discovery of intracellular 2,8 dihydroxyadenine crystals in bovine lymphatic and hepatic cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010012881x ◽

1987 ◽

Vol 45 ◽

pp. 906-907

Author(s):

W. E. Rigsby ◽

D. M. Hinton ◽

V. J. Hurst ◽

P. C. McCaskey

Keyword(s):

Polarized Light ◽

Paraffin Sections ◽

Tissue Samples ◽

Whole Cells ◽

Tissue Sections ◽

Hepatic Cells ◽

Slaughter House ◽

Mammalian Tissues ◽

Carbon Coated ◽

Cellular Components

Crystalline intracellular inclusions are rarely seen in mammalian tissues and are often difficult to positively identify. Lymph node and liver tissue samples were obtained from two cows which had been rejected at the slaughter house due to the abnormal appearance of these organs in the animals. The samples were fixed in formaldehyde and some of the fixed material was embedded in paraffin. Examination of the paraffin sections with polarized light microscopy revealed the presence of numerous crystals in both hepatic and lymph tissue sections. Tissue sections were then deparaffinized in xylene, mounted, carbon coated, and examined in a Phillips 505T SEM equipped with a Tracor Northern X-ray Energy Dispersive Spectroscopy (EDS) system. Crystals were obscured by cellular components and membranes so that EDS spectra were only obtainable from whole cells. Tissue samples which had been fixed but not paraffin-embedded were dehydrated, embedded in Spurrs plastic, and sectioned.

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