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Total Internal Reflection Fluorescence Microscopy
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] |
Total Internal Reflection
Total internal reflection (TIR) is the optical 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 as air, ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Exponential Decay
A quantity is subject to exponential decay if it decreases at a rate proportional to its current value. Symbolically, this process can be expressed by the following differential equation, where is the quantity and (lambda) is a positive rate called the exponential decay constant, disintegration constant, rate constant, or transformation constant: :\frac = -\lambda N. The solution to this equation (see derivation below) is: :N(t) = N_0 e^, where is the quantity at time , is the initial quantity, that is, the quantity at time . Measuring rates of decay Mean lifetime If the decaying quantity, ''N''(''t''), is the number of discrete elements in a certain set, it is possible to compute the average length of time that an element remains in the set. This is called the mean lifetime (or simply the lifetime), where the exponential time constant, \tau, relates to the decay rate constant, λ, in the following way: :\tau = \frac. The mean lifetime can be looked at as a ... [...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), 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 "evanescent": Having Po ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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 ... [...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, 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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Confocal Microscopy
Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser confocal scanning microscopy (LCSM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a 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 concept The principle of co ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Snell's Law
Snell's law (also known as Snell–Descartes law and 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. This law was named after the Dutch astronomer and mathematician Willebrord Snellius (also called Snell). 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. Snell's law states that, for a given pair of media, the ratio of the sines of angle of incidence (\theta_1 ) and angle of refraction (\theta_2 ) is equal to the refractive index of the second medium w.r.t the first (n21) which is equal to the ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Total Internal Reflection Fluorescence Microscopy
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] |
Stray Light
Stray light is light in an optical system, which was not intended in the design. The light may be from the intended source, but follow paths other than intended, or it may be from a source other than the intended source. This light will often set a working limit on the dynamic range of the system; it limits the signal-to-noise ratio or contrast ratio, by limiting how dark the system can be. Ocular straylight is stray light in the human eye. Optical systems Monochromatic light Optical measuring instruments that work with monochromatic light, such as spectrophotometers, define stray light as light in the system at wavelengths (colors) other than the one intended. The stray light level is one of the most critical specifications of an instrument. For instance, intense, narrow absorption bands can easily appear to have a peak absorption less than the true absorption of the sample because the ability of the instrument to measure light transmission through the sample is limited by the st ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Biophysics
Biophysics is an interdisciplinary science that applies approaches and methods traditionally used in physics to study biological phenomena. Biophysics covers all scales of biological organization, from molecular to organismic and populations. Biophysical research shares significant overlap with biochemistry, molecular biology, physical chemistry, physiology, nanotechnology, bioengineering, computational biology, biomechanics, developmental biology and systems biology. The term ''biophysics'' was originally introduced by Karl Pearson in 1892.Roland Glaser. Biophysics: An Introduction'. Springer; 23 April 2012. . The term ''biophysics'' is also regularly used in academia to indicate the study of the physical quantities (e.g. electric current, temperature, stress, entropy) in biological systems. Other biological sciences also perform research on the biophysical properties of living organisms including molecular biology, cell biology, chemical biology, and biochemistry. Overview ... [...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] |
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