Thursday, 19 December 2013

Definitions in Microscopy - For MDS - Oral Pathology students

Azimuth angle. A term used to describe the orientation of an object in a plane such as the specimen plane of the microscope. On a graph with polar coordinates marked off in 360°, the angle subtended between a fixed reference (designated 0°) and a vector rotated about the origin.

Back aperture of the objective lens. An aperture plane of the light microscope located at or near the rear lens element of the objective lens and the site of formation of a diffraction image of the object. Optical devices such as phase plates, DIC prisms, and aperture masks used in forms of interference microscopy are located at or near this location.

Background subtraction. An operation in electronic imaging whereby an image of the featureless background near an object is subtracted from the image of the specimen to remove patterns of irregular illumination and other optical faults such as scratches and dust.

Beam splitter. An optical device for separating an incident beam of light into two or more beams. A prism beam splitter in the trinocular head of a microscope directs the imaging beam to the eyepieces and to the camera simultaneously. A polarizing beam splitter made of a crystalline birefringent material is used to produce linearly polarized light. A dichroic mirror beam splitter reflects excitation wavelengths while transmitting long-wavelength fluorescence emission.

Bertrand lens. A built-in telescope lens located behind the back aperture of the objective lens. When rotated into the optical path, the back aperture and diffraction plane are seen, and other planes that are conjugate to it, while looking in the oculars of the microscope.

Biaxial crystal. A class of birefringent crystals having two optic axes. Mica is an example of this class of crystals.

Bioluminescence. An oxidative process resulting in the release of energy as light emission— for example, firefly luminescence, which requires an enzyme, luciferase, to catalyze a reaction between the substrate luciferin and molecular oxygen in the presence of ATP. 181

Birefringence. The double refraction of light in transparent, molecularly ordered materials caused by the existence of orientation-dependent differences in refractive index. Also refers to the refractive index difference experienced by a transmitted ray through such a material. Incident beams of light on a birefringent specimen are split into O and E rays that can recombine after emergence from the object, giving linearly, elliptically, or circularly polarized light.

Brewster’s angle. The unique angle formed by an incident ray and the perpendicular to a reflective dielectric substance such as water or glass (the transmitting medium) at which the reflected ray is totally linearly polarized.

Bright-field microscopy. A mode of optics employing the basic components of objective and condenser lenses for the examination of amplitude objects such as stained histological specimens.

Brightness. A qualitative expression for the intensity of light.

Chromatic aberration. An aberration of lenses, whereby light waves of different wavelength are brought to focus at different locations along the optic axis. In the typical case of a simple thin lens, the focal length is shorter for blue wavelengths than it is for red ones. 5


Coherent light. A beam of light defined by waves vibrating in the same phase, although not necessarily in the same plane of vibration. To maintain the same phase over long distances, coherent waves must be monochromatic (have the same wavelength). Laser light is monochromatic, linearly polarized, and highly coherent. 

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