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Dark Field Microscopy
Dark-field microscopy (also called dark-ground microscopy) describes microscopy methods, in both light and electron microscopy, which exclude the unscattered beam from the image. As a result, the field around the specimen (i.e., where there is no specimen to scatter the beam) is generally dark. In optical microscopes a darkfield condenser lens must be used, which directs a cone of light away from the objective lens. To maximize the scattered light-gathering power of the objective lens, oil immersion is used and the numerical aperture (NA) of the objective lens must be less than 1.0. Objective lenses with a higher NA can be used but only if they have an adjustable diaphragm, which reduces the NA. Often these objective lenses have a NA that is variable from 0.7 to 1.25. Light microscopy applications In optical microscopy, dark-field describes an illumination technique used to enhance the contrast in unstained samples. It works by illuminating the sample with light that will ...
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Red Blood Cells By Darkfield Microscopy
Red is the color at the long wavelength end of the visible spectrum of light, next to orange and opposite violet. It has a dominant wavelength of approximately 625–740 nanometres. It is a primary color in the RGB color model and a secondary color (made from magenta and yellow) in the CMYK color model, and is the complementary color of cyan. Reds range from the brilliant yellow-tinged scarlet and vermillion to bluish-red crimson, and vary in shade from the pale red pink to the dark red burgundy. Red pigment made from ochre was one of the first colors used in prehistoric art. The Ancient Egyptians and Mayans colored their faces red in ceremonies; Roman generals had their bodies colored red to celebrate victories. It was also an important color in China, where it was used to color early pottery and later the gates and walls of palaces. In the Renaissance, the brilliant red costumes for the nobility and wealthy were dyed with kermes and cochineal. The 19th century bro ...
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Staining (biology)
Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Stains may be used to define biological tissues (highlighting, for example, muscle fibers or connective tissue), cell populations (classifying different blood cells), or organelles within individual cells. In biochemistry, it involves adding a class-specific ( DNA, proteins, lipids, carbohydrates) dye to a substrate to qualify or quantify the presence of a specific compound. Staining and fluorescent tagging can serve similar purposes. Biological staining is also used to mark cells in flow cytometry, and to flag proteins or nucleic acids in gel electrophoresis. Light microscopes are used for viewing s ...
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Hyperspectral Imaging
Hyperspectral imaging collects and processes information from across the electromagnetic spectrum. The goal of hyperspectral imaging is to obtain the spectrum for each pixel in the image of a scene, with the purpose of finding objects, identifying materials, or detecting processes. There are three general branches of spectral imagers. There are push broom scanners and the related whisk broom scanners (spatial scanning), which read images over time, band sequential scanners (spectral scanning), which acquire images of an area at different wavelengths, and snapshot hyperspectral imaging, which uses a staring array to generate an image in an instant. Whereas the human eye sees color of visible light in mostly three bands (long wavelengths - perceived as red, medium wavelengths - perceived as green, and short wavelengths - perceived as blue), spectral imaging divides the spectrum into many more bands. This technique of dividing images into bands can be extended beyond the visible. ...
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Optical Mouse
An optical mouse is a computer mouse which uses a light source, typically a light-emitting diode (LED), and a light detector, such as an array of photodiodes, to detect movement relative to a surface. Variations of the optical mouse have largely replaced the older mechanical mouse design, which uses moving parts to sense motion. The earliest optical mice detected movement on pre-printed mousepad surfaces. Modern optical mice work on most opaque diffusely reflective surfaces like paper, but most of them do not work properly on specularly reflective surfaces like polished stone or transparent surfaces like glass. Optical mice that use dark field illumination can function reliably even on such surfaces. Mechanical mice Though not commonly referred to as optical mice, nearly all mechanical mice tracked movement using LEDs and photodiodes to detect when beams of infrared light did and didn't pass through holes in a pair of incremental rotary encoder wheels (one for left/rig ...
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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 ...
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Phase-contrast Microscopy
__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 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 cellular structures that are invisible with a bright-field microscope, as exemp ...
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Polarization (waves)
Polarization ( also polarisation) is a property applying to transverse waves that specifies the geometrical orientation of the oscillations. In a transverse wave, the direction of the oscillation is perpendicular to the direction of motion of the wave. A simple example of a polarized transverse wave is vibrations traveling along a taut string ''(see image)''; for example, in a musical instrument like a guitar string. Depending on how the string is plucked, the vibrations can be in a vertical direction, horizontal direction, or at any angle perpendicular to the string. In contrast, in longitudinal waves, such as sound waves in a liquid or gas, the displacement of the particles in the oscillation is always in the direction of propagation, so these waves do not exhibit polarization. Transverse waves that exhibit polarization include electromagnetic waves such as light and radio waves, gravitational waves, and transverse sound waves (shear waves) in solids. An electromagnetic ...
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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 ...
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Attenuation Coefficient
The linear attenuation coefficient, attenuation coefficient, or narrow-beam attenuation coefficient characterizes how easily a volume of material can be penetrated by a beam of light, sound, particles, or other energy or matter. A coefficient value that is large represents a beam becoming 'attenuated' as it passes through a given medium, while a small value represents that the medium had little effect on loss. The SI unit of attenuation coefficient is the reciprocal metre (m−1). Extinction coefficient is another term for this quantity, often used in meteorology and climatology. Most commonly, the quantity measures the exponential decay of intensity, that is, the value of downward ''e''-folding distance of the original intensity as the energy of the intensity passes through a unit (''e.g.'' one meter) thickness of material, so that an attenuation coefficient of 1 m−1 means that after passing through 1 metre, the radiation will be reduced by a factor of '' e'', and for material ...
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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 lig ...
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Scattered Radiation
Scattering is a term used in physics to describe a wide range of physical processes where moving particles or radiation of some form, such as light or sound, are forced to deviate from a straight trajectory by localized non-uniformities (including particles and radiation) in the medium through which they pass. In conventional use, this also includes deviation of reflected radiation from the angle predicted by the law of reflection. Reflections of radiation that undergo scattering are often called ''diffuse reflections'' and unscattered reflections are called ''specular'' (mirror-like) reflections. Originally, the term was confined to light scattering (going back at least as far as Isaac Newton in the 17th century). As more "ray"-like phenomena were discovered, the idea of scattering was extended to them, so that William Herschel could refer to the scattering of "heat rays" (not then recognized as electromagnetic in nature) in 1800. John Tyndall, a pioneer in light scattering researc ...
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Tissue Paper
Tissue paper or simply tissue is a lightweight paper or, light crêpe paper. Tissue can be made from recycled paper pulp on a paper machine. Tissue paper is very versatile, and different kinds of tissue are made to best serve these purposes, which are hygienic tissue paper, facial tissues, paper towels, as packing material, among other (sometimes creative) uses. The use of tissue paper is common in developed nations, around 21 million tonnes in North America and 6 million in Europe, and is growing due to urbanization. As a result, the industry has often been scrutinized for deforestation. However, more companies are presently using more recycled fibres in tissue paper. Properties The key properties of tissues are absorbency, basis weight, thickness, bulk (specific volume), brightness, stretch, appearance and comfort. Production Tissue paper is produced on a paper machine that has a single large steam heated drying cylinder ( Yankee dryer) fitted with a hot air hood. The ...
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