Bright Field Microscopy
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Bright Field Microscopy
Bright-field microscopy (BF) is the simplest of all the optical microscopy illumination techniques. Sample illumination is transmitted (i.e., illuminated from below and observed from above) white light, and contrast in the sample is caused by attenuation of the transmitted light in dense areas of the sample. Bright-field microscopy is the simplest of a range of techniques used for illumination of samples in light microscopes, and its simplicity makes it a popular technique. The typical appearance of a bright-field microscopy image is a dark sample on a bright background, hence the name. Light path The light path of a bright-field microscope is extremely simple, no additional components are required beyond the normal light-microscope setup. The light path therefore consists of: * a transillumination light source, commonly a halogen lamp in the microscope stand; * a condenser lens, which focuses light from the light source onto the sample; * an objective lens, which collects light ...
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Polarized Light Microscopy
Polarized light microscopy can mean any of a number of optical microscopy techniques involving polarized light. Simple techniques include illumination of the sample with polarized light. Directly transmitted light can, optionally, be blocked with a polariser orientated at 90 degrees to the illumination. More complex microscopy techniques which take advantage of polarized light include differential interference contrast microscopy and interference reflection microscopy. Scientists will often use a device called a polarizing plate to convert natural light into polarized light. These illumination techniques are most commonly used on birefringent samples where the polarized light interacts strongly with the sample and so generating contrast with the background. Polarized light microscopy is used extensively in optical mineralogy. History Although the invention of the polarizing microscope is typically attributed to David Brewster around 1815, Brewster clearly acknowledges the prio ...
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Iris Diaphragm
In optics, a diaphragm is a thin opaque structure with an opening (aperture) at its center. The role of the diaphragm is to ''stop'' the passage of light, except for the light passing through the ''aperture''. Thus it is also called a stop (an aperture stop, if it limits the brightness of light reaching the focal plane, or a field stop or flare stop for other uses of diaphragms in lenses). The diaphragm is placed in the light path of a lens or objective, and the size of the aperture regulates the amount of light that passes through the lens. The centre of the diaphragm's aperture coincides with the optical axis of the lens system. Most modern cameras use a type of adjustable diaphragm known as an iris diaphragm, and often referred to simply as an iris. See the articles on aperture and f-number for the photographic effect and system of quantification of varying the opening in the diaphragm. Iris diaphragms versus other types A natural optical system that has a diaphragm an ...
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Chara (alga)
''Chara'' is a genus of charophyte green algae in the family Characeae. They are multicellular and superficially resemble land plants because of stem-like and leaf-like structures. They are found in freshwater, particularly in limestone areas throughout the northern temperate zone, where they grow submerged, attached to the muddy bottom. They prefer less oxygenated and hard water and are not found in waters where mosquito larvae are present. They are covered with calcium carbonate deposits and are commonly known as stoneworts. Cyanobacteria have been found growing as epiphytes on the surfaces of ''Chara'', where they may be involved in fixing nitrogen, which is important to plant nutrition. Structure The branching system of ''Chara'' species is complex with branches derived from apical cells which cut off segments at the base to form nodal and internodal cells alternately.Round, F.E. 1965.''The Biology of the algae.'' Ernest Arnold. The main axes bear whorls of branches in a ...
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Cytoplasmic Streaming
Cytoplasmic streaming, also called protoplasmic streaming and cyclosis, is the flow of the cytoplasm inside the cell, driven by forces from the cytoskeleton. It is likely that its function is, at least in part, to speed up the transport of molecules and organelles around the cell. It is usually observed in large plant and animal cells, greater than approximately 0.1 mm. In smaller cells, the diffusion of molecules is more rapid, but diffusion slows as the size of the cell increases, so larger cells may need cytoplasmic streaming for efficient function. The green alga genus ''Chara'' possesses some very large cells, up to 10 cm in length, and cytoplasmic streaming has been studied in these large cells. Cytoplasmic streaming is strongly dependent upon intracellular pH and temperature. It has been observed that the effect of temperature on cytoplasmic streaming created linear variance and dependence at different high temperatures in comparison to low temperatures. This ...
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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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Visible Light
Light or visible light is electromagnetic radiation that can be perceived by the human eye. Visible light is usually defined as having wavelengths in the range of 400–700 nanometres (nm), corresponding to frequencies of 750–420 terahertz, between the infrared (with longer wavelengths) and the ultraviolet (with shorter wavelengths). In physics, the term "light" may refer more broadly to electromagnetic radiation of any wavelength, whether visible or not. In this sense, gamma rays, X-rays, microwaves and radio waves are also light. The primary properties of light are intensity, propagation direction, frequency or wavelength spectrum and polarization. Its speed in a vacuum, 299 792 458 metres a second (m/s), is one of the fundamental constants of nature. Like all types of electromagnetic radiation, visible light propagates by massless elementary particles called photons that represents the quanta of electromagnetic field, and can be analyzed as both waves and ...
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Wavelength
In physics, the wavelength is the spatial period of a periodic wave—the distance over which the wave's shape repeats. It is the distance between consecutive corresponding points of the same phase on the wave, such as two adjacent crests, troughs, or zero crossings, and is a characteristic of both traveling waves and standing waves, as well as other spatial wave patterns. The inverse of the wavelength is called the spatial frequency. Wavelength is commonly designated by the Greek letter ''lambda'' (λ). The term ''wavelength'' is also sometimes applied to modulated waves, and to the sinusoidal envelopes of modulated waves or waves formed by interference of several sinusoids. Assuming a sinusoidal wave moving at a fixed wave speed, wavelength is inversely proportional to frequency of the wave: waves with higher frequencies have shorter wavelengths, and lower frequencies have longer wavelengths. Wavelength depends on the medium (for example, vacuum, air, or water) that a wav ...
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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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Magnification
Magnification is the process of enlarging the apparent size, not physical size, of something. This enlargement is quantified by a calculated number also called "magnification". When this number is less than one, it refers to a reduction in size, sometimes called ''minification'' or ''de-magnification''. Typically, magnification is related to scaling up visuals or images to be able to see more detail, increasing resolution, using microscope, printing techniques, or digital processing. In all cases, the magnification of the image does not change the perspective of the image. Examples of magnification Some optical instruments provide visual aid by magnifying small or distant subjects. * A magnifying glass, which uses a positive (convex) lens to make things look bigger by allowing the user to hold them closer to their eye. * A telescope, which uses its large objective lens or primary mirror to create an image of a distant object and then allows the user to examine the image clo ...
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Interference (wave Propagation)
In physics, interference is a phenomenon in which two waves combine by adding their displacement together at every single point in space and time, to form a resultant wave of greater, lower, or the same amplitude. Constructive and destructive interference result from the interaction of waves that are correlated or coherent with each other, either because they come from the same source or because they have the same or nearly the same frequency. Interference effects can be observed with all types of waves, for example, light, radio, acoustic, surface water waves, gravity waves, or matter waves. Etymology The word ''interference'' is derived from the Latin words ''inter'' which means "between" and ''fere'' which means "hit or strike", and was coined by Thomas Young in 1801. Mechanisms The principle of superposition of waves states that when two or more propagating waves of the same type are incident on the same point, the resultant amplitude at that point is equal to th ...
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Phase-contrast
__NOTOC__ Phase-contrast microscopy (PCM) is an optical microscopy technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image. Phase shifts themselves are invisible, but become visible when shown as brightness variations. When light waves travel through a medium other than a vacuum, interaction with the medium causes the wave amplitude and phase (waves), phase to change in a manner dependent on properties of the medium. Changes in amplitude (brightness) arise from the scattering and absorption of light, which is often wavelength-dependent and may give rise to colors. Photographic equipment and the human eye are only sensitive to amplitude variations. Without special arrangements, phase changes are therefore invisible. Yet, phase changes often convey important information. Phase-contrast microscopy is particularly important in biology. It reveals many cell (biology), cellular structures that are invisible with a brigh ...
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