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Water Immersion Objective
In light microscopy, a water immersion objective is a specially designed objective lens used to increase the resolution of the microscope. This is achieved by immersing both the lens and the specimen in water which has a higher refractive index than air, thereby increasing the numerical aperture of the objective lens. Applications Water immersion objectives are used not only at very large magnifications that require high resolving power, but also of moderate power as there are water immersion objectives as low as 4X. Objectives with high power magnification have short focal lengths, facilitating the use of water. The water is applied to the specimen (conventional microscope), and the stage is raised, immersing the objective in water. Sometimes with water dipping objectives, the objective is directly immersed in the solution of water which contains the specimens to look at. Electrophoretic preparations used in the case of comet assay can benefit from the use of water objectives. T ...
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Light Microscopy
Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye (objects that are not within the resolution range of the normal eye). There are three well-known branches of microscopy: optical microscope, optical, electron microscope, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy. Optical microscopy and electron microscopy involve the diffraction, reflection (physics), reflection, or refraction of electromagnetic radiation/electron beams interacting with the Laboratory specimen, specimen, and the collection of the scattered radiation or another signal in order to create an image. This process may be carried out by wide-field irradiation of the sample (for example standard light microscopy and transmission electron microscope, transmission electron microscopy) or by scanning a fine beam over the sample (for example confocal laser scanning microscopy and scanning electro ...
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Objective (optics)
In optical engineering, the objective is the optical element that gathers light from the object being observed and focuses the light rays to produce a real image. Objectives can be a single lens or mirror, or combinations of several optical elements. They are used in microscopes, binoculars, telescopes, cameras, slide projectors, CD players and many other optical instruments. Objectives are also called object lenses, object glasses, or objective glasses. Microscope objectives The objective lens of a microscope is the one at the bottom near the sample. At its simplest, it is a very high-powered magnifying glass, with very short focal length. This is brought very close to the specimen being examined so that the light from the specimen comes to a focus inside the microscope tube. The objective itself is usually a cylinder containing one or more lenses that are typically made of glass; its function is to collect light from the sample. Magnification One of the most important prope ...
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Optical Resolution
Optical resolution describes the ability of an imaging system to resolve detail, in the object that is being imaged. An imaging system may have many individual components, including one or more lenses, and/or recording and display components. Each of these contributes (given suitable design, and adequate alignment) to the optical resolution of the system; the environment in which the imaging is done often is a further important factor. Lateral resolution Resolution depends on the distance between two distinguishable radiating points. The sections below describe the theoretical estimates of resolution, but the real values may differ. The results below are based on mathematical models of Airy discs, which assumes an adequate level of contrast. In low-contrast systems, the resolution may be much lower than predicted by the theory outlined below. Real optical systems are complex, and practical difficulties often increase the distance between distinguishable point sources. The ...
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Refractive Index
In optics, the refractive index (or refraction index) of an optical medium is a dimensionless number that gives the indication of the light bending ability of that medium. The refractive index determines how much the path of light is bent, or refracted, when entering a material. This is described by Snell's law of refraction, , where ''θ''1 and ''θ''2 are the angle of incidence and angle of refraction, respectively, of a ray crossing the interface between two media with refractive indices ''n''1 and ''n''2. The refractive indices also determine the amount of light that is reflected when reaching the interface, as well as the critical angle for total internal reflection, their intensity ( Fresnel's equations) and Brewster's angle. The refractive index can be seen as the factor by which the speed and the wavelength of the radiation are reduced with respect to their vacuum values: the speed of light in a medium is , and similarly the wavelength in that medium is , where ''Π...
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Numerical Aperture
In optics, the numerical aperture (NA) of an optical system is a dimensionless number that characterizes the range of angles over which the system can accept or emit light. By incorporating index of refraction in its definition, NA has the property that it is constant for a beam as it goes from one material to another, provided there is no refractive power at the interface. The exact definition of the term varies slightly between different areas of optics. Numerical aperture is commonly used in microscopy to describe the acceptance cone of an objective (and hence its light-gathering ability and resolution), and in fiber optics, in which it describes the range of angles within which light that is incident on the fiber will be transmitted along it. General optics In most areas of optics, and especially in microscopy, the numerical aperture of an optical system such as an objective lens is defined by :\mathrm = n \sin \theta, where is the index of refraction of the medium i ...
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Focal Length
The focal length of an optical system is a measure of how strongly the system converges or diverges light; it is the inverse of the system's optical power. A positive focal length indicates that a system converges light, while a negative focal length indicates that the system diverges light. A system with a shorter focal length bends the rays more sharply, bringing them to a focus in a shorter distance or diverging them more quickly. For the special case of a thin lens in air, a positive focal length is the distance over which initially collimated (parallel) rays are brought to a focus, or alternatively a negative focal length indicates how far in front of the lens a point source must be located to form a collimated beam. For more general optical systems, the focal length has no intuitive meaning; it is simply the inverse of the system's optical power. In most photography and all telescopy, where the subject is essentially infinitely far away, longer focal length (lower opti ...
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Comet Assay
A comet is an icy, small Solar System body that, when passing close to the Sun, warms and begins to release gases, a process that is called outgassing. This produces a visible atmosphere or coma, and sometimes also a tail. These phenomena are due to the effects of solar radiation and the solar wind acting upon the nucleus of the comet. Comet nuclei range from a few hundred meters to tens of kilometers across and are composed of loose collections of ice, dust, and small rocky particles. The coma may be up to 15 times Earth's diameter, while the tail may stretch beyond one astronomical unit. If sufficiently bright, a comet may be seen from Earth without the aid of a telescope and may subtend an arc of 30° (60 Moons) across the sky. Comets have been observed and recorded since ancient times by many cultures and religions. Comets usually have highly eccentric elliptical orbits, and they have a wide range of orbital periods, ranging from several years to potentially several milli ...
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Numerical Apertures
In optics, the numerical aperture (NA) of an optical system is a dimensionless number that characterizes the range of angles over which the system can accept or emit light. By incorporating index of refraction in its definition, NA has the property that it is constant for a beam as it goes from one material to another, provided there is no refractive power at the interface. The exact definition of the term varies slightly between different areas of optics. Numerical aperture is commonly used in microscopy to describe the acceptance cone of an objective (and hence its light-gathering ability and resolution), and in fiber optics, in which it describes the range of angles within which light that is incident on the fiber will be transmitted along it. General optics In most areas of optics, and especially in microscopy, the numerical aperture of an optical system such as an objective lens is defined by :\mathrm = n \sin \theta, where is the index of refraction of the medium in whi ...
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Oil Immersion Objective
In light microscopy, oil immersion is a technique used to increase the resolving power of a microscope. This is achieved by immersing both the objective lens and the specimen in a transparent oil of high refractive index, thereby increasing the numerical aperture of the objective lens. Without oil, light waves reflect off the slide specimen through the glass cover slip, through the air, and into the microscope lens (see the colored figure to the right). Unless a wave comes out at a 90-degree angle, it bends when it hits a new substance, the amount of bend depending on the angle. This distorts the image. Air has a very different index of refraction from glass, making for a larger bend compared to oil, which has an index more similar to glass. Specially manufactured oil can have nearly exactly the same refractive index as glass, making an oil immersed lens nearly as effective as having entirely glass to the sample (which would be impractical). Immersion oils are transparent oils t ...
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Microscopy
Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye (objects that are not within the resolution range of the normal eye). There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy. Optical microscopy and electron microscopy involve the diffraction, reflection, or refraction of electromagnetic radiation/electron beams interacting with the specimen, and the collection of the scattered radiation or another signal in order to create an image. This process may be carried out by wide-field irradiation of the sample (for example standard light microscopy and transmission electron microscopy) or by scanning a fine beam over the sample (for example confocal laser scanning microscopy and scanning electron microscopy). Scanning probe microscopy involves the interaction of a scanning probe with the surface of the objec ...
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Optical Microscope
The optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century. Basic optical microscopes can be very simple, although many complex designs aim to improve resolution and sample contrast. The object is placed on a stage and may be directly viewed through one or two eyepieces on the microscope. In high-power microscopes, both eyepieces typically show the same image, but with a stereo microscope, slightly different images are used to create a 3-D effect. A camera is typically used to capture the image (micrograph). The sample can be lit in a variety of ways. Transparent objects can be lit from below and solid objects can be lit with light coming through ( bright field) or around (dark field) the objective lens. Polarised ...
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Index-matching Material
In optics, an index-matching material is a substance, usually a liquid, cement (adhesive), or gel, which has an index of refraction that closely approximates that of another object (such as a lens, material, fiber-optic, etc.). When two substances with the same index are in contact, light passes from one to the other with neither reflection nor refraction. As such, they are used for various purposes in science, engineering, and art. For example, in a popular home experiment, a glass rod is made almost invisible by immersing it in an index-matched transparent fluid such as mineral spirits. In microscopy In light microscopy, oil immersion is a technique used to increase the resolution of a microscope. This is achieved by immersing both the objective lens and the specimen in a transparent oil of high refractive index, thereby increasing the numerical aperture of the objective lens. Immersion oils are transparent oils that have specific optical and viscosity characteristics necessar ...
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