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Total Internal Reflection Fluorescence Microscope
A total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nanometers can be observed. TIRFM is an imaging modality which uses the excitation of fluorescent cells in a thin optical specimen section that is supported on a glass slide. The technique is based on the principle that when excitation light is totally internally reflected in a transparent solid coverglass at its interface with a liquid medium, an electromagnetic field, also known as an evanescent wave, is generated at the solid-liquid interface with the same frequency as the excitation light. The intensity of the evanescent wave exponentially decays with distance from the surface of the solid so that only fluorescent molecules within a few hundred nanometers of the solid are efficiently excited. Two-dimensional images of the fluorescence can then be obtained, although there are also mechanisms in which three-dimensional information on the ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Evanescent Field
In electromagnetics, an evanescent field, or evanescent wave, is an oscillating electric and/or magnetic field that does not propagate as an electromagnetic wave but whose energy is spatially concentrated in the vicinity of the source (oscillating charges and currents). Even when there is a propagating electromagnetic wave produced (e.g., by a transmitting Antenna (radio), antenna), one can still identify as an evanescent field the component of the electric or magnetic field that cannot be attributed to the propagating wave observed at a distance of many wavelengths (such as the far field of a transmitting antenna). A hallmark of an evanescent field is that there is no net energy flow in that region. Since the net flow of electromagnetic energy is given by the average Poynting vector, this means that the Poynting vector in these regions, as averaged over a complete oscillation cycle, is zero. Use of the term In many cases one cannot simply say that a field is or is not "evane ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Single-molecule Experiment
A single-molecule experiment is an experiment that investigates the properties of individual molecules. Single-molecule studies may be contrasted with measurements on an ensemble or bulk collection of molecules, where the individual behavior of molecules cannot be distinguished, and only average characteristics can be measured. Since many measurement techniques in biology, chemistry, and physics are not sensitive enough to observe single molecules, single-molecule fluorescence techniques (that have emerged since the 1990s for probing various processes on the level of individual molecules) caused a lot of excitement, since these supplied many new details on the measured processes that were not accessible in the past. Indeed, since the 1990s, many techniques for probing individual molecules have been developed. The first single-molecule experiments were patch clamp experiments performed in the 1970s, but these were limited to studying ion channels. Today, systems investigated using s ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Fluorescence Microscope
A fluorescence microscope is an optical microscope that uses fluorescence instead of, or in addition to, scattering, reflection, and attenuation or absorption, to study the properties of organic or inorganic substances. A fluorescence microscope is any microscope that uses fluorescence to generate an image, whether it is a simple setup like an epifluorescence microscope or a more complicated design such as a confocal microscope, which uses optical sectioning to get better resolution of the fluorescence image. Principle The specimen is illuminated with light of a specific wavelength (or wavelengths) which is absorbed by the fluorophores, causing them to emit light of longer wavelengths (i.e., of a different color than the absorbed light). The illumination light is separated from the much weaker emitted fluorescence through the use of a spectral emission filter. Typical components of a fluorescence microscope are a light source (xenon arc lamp or mercury-vapor lamp are com ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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, 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 (e.g., a flat 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 (optics), objective (and hence its light-gathering ability and Optical resolution, 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 \t ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Objective (optics)
In optical engineering, an objective is an optical element that gathers light from an object being observed and Focus (optics), focuses the ray (optics), light rays from it to produce a real image of the object. Objectives can be a single Lens (optics), 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 fr ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Visible Light
Light, visible light, or visible radiation is electromagnetic radiation that can be perceived by the human eye. Visible light spans the visible spectrum and is usually defined as having wavelengths in the range of 400–700 nanometres (nm), corresponding to frequencies of 750–420 terahertz. The visible band sits adjacent to the infrared (with longer wavelengths and lower frequencies) and the ultraviolet (with shorter wavelengths and higher frequencies), called collectively '' optical radiation''. 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 vacuum, , is one of the fundamental constants of nature. All electromagnetic radiation exhibits some properties of both particles and waves ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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, 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 (e.g., a flat 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 (optics), objective (and hence its light-gathering ability and Optical resolution, 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 \t ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Confocal Microscopy
Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast (vision), contrast of a micrograph by means of using a Spatial filter, spatial pinhole to block out-of-focus light in image formation. Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures (a process known as optical sectioning) within an object. This technique is used extensively in the scientific and industrial communities and typical applications are in life sciences, semiconductor inspection and materials science. Light travels through the sample under a conventional microscope as far into the specimen as it can penetrate, while a confocal microscope only focuses a smaller beam of light at one narrow depth level at a time. The CLSM achieves a controlled and highly limited depth of field. Basic c ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Snell's Law
Snell's law (also known as the Snell–Descartes law, the ibn-Sahl law, and the law of refraction) is a formula used to describe the relationship between the angles of incidence and refraction, when referring to light or other waves passing through a boundary between two different isotropic media, such as water, glass, or air. In optics, the law is used in ray tracing to compute the angles of incidence or refraction, and in experimental optics to find the refractive index of a material. The law is also satisfied in meta-materials, which allow light to be bent "backward" at a negative angle of refraction with a negative refractive index. The law states that, for a given pair of media, the ratio of the sines of angle of incidence \left(\theta_1 \right) and angle of refraction \left(\theta_2\right) is equal to the refractive index of the second medium with regard to the first (n_) which is equal to the ratio of the refractive indices \left(\tfrac\right) of the two media, or e ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Total Internal Reflection
In physics, total internal reflection (TIR) is the phenomenon in which waves arriving at the interface (boundary) from one medium to another (e.g., from water to air) are not refracted into the second ("external") medium, but completely reflected back into the first ("internal") medium. It occurs when the second medium has a higher wave speed (i.e., lower refractive index) than the first, and the waves are incident at a sufficiently oblique angle on the interface. For example, the water-to-air surface in a typical fish tank, when viewed obliquely from below, reflects the underwater scene like a mirror with no loss of brightness (Fig.1). TIR occurs not only with electromagnetic waves such as light and microwaves, but also with other types of waves, including sound and water waves. If the waves are capable of forming a narrow beam (Fig.2), the reflection tends to be described in terms of " rays" rather than waves; in a medium whose properties are independent of direction, such ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Tirfm
A total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nanometers can be observed. TIRFM is an imaging modality which uses the excitation of fluorescent cells in a thin optical specimen section that is supported on a glass slide. The technique is based on the principle that when excitation light is totally internally reflected in a transparent solid coverglass at its interface with a liquid medium, an electromagnetic field, also known as an evanescent wave, is generated at the solid-liquid interface with the same frequency as the excitation light. The intensity of the evanescent wave exponentially decays with distance from the surface of the solid so that only fluorescent molecules within a few hundred nanometers of the solid are efficiently excited. Two-dimensional images of the fluorescence can then be obtained, although there are also mechanisms in which three-dimensional information on the ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Collimated Beam
A collimated beam of light or other electromagnetic radiation has parallel rays, and therefore will spread minimally as it propagates. A laser beam is an archetypical example. A perfectly collimated light beam, with no divergence, would not disperse with distance. However, diffraction prevents the creation of any such beam. Light can be approximately collimated by a number of processes, for instance by means of a collimator. Perfectly collimated light is sometimes said to be ''focused at infinity''. Thus, as the distance from a point source increases, the spherical wavefronts become flatter and closer to plane waves, which are perfectly collimated. Other forms of electromagnetic radiation can also be collimated. In radiology, X-rays are collimated to reduce the volume of the patient's tissue that is irradiated, and to remove stray photons that reduce the quality of the x-ray image ("film fog"). In scintigraphy, a gamma ray collimator is used in front of a detector to allow only ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
