Viewing the Specimen Using Reflected-Light Microscopy

2010 ◽  
pp. 89-114

Abstract The analysis of composite materials using optical microscopy is a process that can be made easy and efficient with only a few contrast methods and preparation techniques. This chapter is intended to provide information that will help an investigator select the appropriate microscopy technique for the specific analysis objectives with a given composite material. The chapter opens with a discussion of macrophotography and microscope alignment, and then goes on to describe various illumination techniques that are useful for specific analysis requirements. These techniques include bright-field illumination, dark-field illumination, polarized-light microscopy, interference and contrast microscopy, and fluorescence microscopy. The chapter also provides a discussion of sample preparation materials such as dyes, etchants, and stains for the analysis of composite materials using optical microscopy.

1984 ◽  
pp. 267-333

Abstract This chapter discusses the tools and techniques of light microscopy and how they are used in the study of materials. It reviews the basic physics of light, the inner workings of light microscopes, and the relationship between resolution and depth of field. It explains the difference between amplitude and optical-phase features and how they are revealed using appropriate illumination methods. It compares images obtained using bright field and dark field illumination, polarized and cross-polarized light, and interference-contrast techniques. It also discusses the use of photometers, provides best practices and recommendations for photographing structures and features of interest, and describes the capabilities of hot-stage and hot-cell microscopes.


2020 ◽  
Vol 64 (3) ◽  
Author(s):  
Eva Jennische ◽  
Stefan Lange ◽  
Ragnar Hultborn

A simple dark field microscopy technique was used for visualization of blood vessels in normal human renal tissues and carcinoma. Phase contrast condenser ring apt for high power objectives was combined with a 10x objective in order to create a dark field illumination of the specimens examined. The endothelial lining of the vessels had been stained by using CD31 monoclonal antibodies combined with conventional peroxidase immunohistochemistry. The final DAB addition used for this technique induced an intense light scatter in the dark field microscope. This scattered light originating from the endothelial lining made the walls of the bright vessels easily detectable from the dark background.


Author(s):  
Kannan M. Krishnan

Propagation of light is described as the simple harmonic motion of transverse waves. Combining waves that propagate on orthogonal planes give rise to linear, elliptical, or spherical polarization, depending on their amplitudes and phase differences. Classical experiments of Huygens and Young demonstrated the principle of optical interference and diffraction. Generalization of Fraunhofer diffraction to scattering by a three-dimensional arrangement of atoms in crystals forms the basis of diffraction methods. Fresnel diffraction finds application in the design of zone plates for X-ray microscopy. Optical microscopy, with resolution given by the Rayleigh criterion to be approximately half the wavelength, works best when tailored to the optimal characteristics of the human eye (λ = 550 nm). Lenses suffer from spherical and chromatic aberrations, and astigmatism. Optical microscopes operate in bright-field, oblique, and dark-field imaging conditions, produce interference contrast, and can image with polarized light. Variants include confocal scanning optical microscopy (CSOM). Metallography, widely used to characterize microstructures, requires polished or chemically etched surfaces to provide optimal contrast. Finally, the polarization state of light reflected from the surface of a specimen is utilized in ellipsometry to obtain details of the optical properties and thickness of thin film materials.


IAWA Journal ◽  
2020 ◽  
Vol 41 (4) ◽  
pp. 478-489 ◽  
Author(s):  
Valentina Zemke ◽  
Volker Haag ◽  
Gerald Koch

Abstract The present study focusses on the application of 3D-reflected light microscopy (3D-RLM) for the wood anatomical identification of charcoal specimens produced from domestic and tropical timbers. This special microscopic technique offers a detailed investigation of anatomical features in charcoal directly compared with the quality of field emission scanning electron microscopy (FESEM). The advantages of using the 3D-RLM technology are that fresh fracture planes of charcoal can be directly observed under the microscope without further preparation or surface treatment. Furthermore, the 3D-technique with integrated polarized light illumination creates high-contrast images of uneven and black charcoal surfaces. Important diagnostic structural features such as septate fibres and intercellular canals can be clearly detected and intervessel pits are directly measured. The comparison of the microscopic analyses reveals that 3D-reflected light microscopy (3D-RLM) provides an effective alternative technique to conventional field emission scanning electron microscopy for the identification of carbonized wood.


2010 ◽  
pp. 159-175

Abstract The formation of microcracks in composite materials may arise from static-, dynamic-, impact-, or fatigue-loading situations and also by temperature changes or thermal cycles. This chapter discusses the processes involved in the various methods for the microcrack analysis of composite materials, namely bright-field analysis, polarized-light analysis, contrast dyes analysis, and dark-field analysis. The analysis of microcracked composites using epi-fluorescence is also covered. In addition, the chapter describes the procedures for the determination and recording of microcracks in composite materials.


Author(s):  
A. G. Anisovich

The optical effects which appeared on internal defects of optically transparent materials by use of various methods of optical staining, i. e. dark-field illumination and polarized light were researched. It was shown that methods of optical staining support to determine spherical defects under a surface of optically transparent materials. Formation of optical effects on materials defects in dark background are partially determined by design features of microscope objective and it was found out. It was defined that the investigation using polarized light the image formation of spherical defects occurs similar to uniaxial crystal. 


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