Super-resolution Optical Fluctuation Imaging
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Super-resolution Optical Fluctuation Imaging
Super-resolution optical fluctuation imaging (SOFI) is a post-processing method for the calculation of super-resolved images from recorded image time series that is based on the temporal correlations of independently fluctuating fluorescent emitters. SOFI has been developed for super-resolution of biological specimen that are labelled with independently fluctuating fluorescent emitters (organic dyes, fluorescent proteins). In comparison to other super-resolution microscopy techniques such as STORM or PALM that rely on single-molecule localization and hence only allow one active molecule per diffraction-limited area (DLA) and timepoint, SOFI does not necessitate a controlled photoswitching and/ or photoactivation as well as long imaging times. Nevertheless, it still requires fluorophores that are cycling through two distinguishable states, either real on-/off-states or states with different fluorescence intensities. In mathematical terms SOFI-imaging relies on the calculation of cumu ...
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Green Fluorescent Protein
The green fluorescent protein (GFP) is a protein that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. The label ''GFP'' traditionally refers to the protein first isolated from the jellyfish ''Aequorea victoria'' and is sometimes called ''avGFP''. However, GFPs have been found in other organisms including corals, sea anemones, zoanithids, copepods and lancelets. The GFP from ''A. victoria'' has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum. The fluorescence quantum yield (QY) of GFP is 0.79. The GFP from the sea pansy (''Renilla reniformis'') has a single major excitation peak at 498 nm. GFP makes for an excellent tool in many forms of biology due to its ability to form an internal chromophore without requiring any accessory cofactors, gene products, or enzymes / substrates other than mo ...
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Super-resolution Microscopy
Super-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, which is due to the diffraction of light. Super-resolution imaging techniques rely on the near-field (photon-tunneling microscopy as well as those that utilize the Pendry Superlens and near field scanning optical microscopy) or on the far-field. Among techniques that rely on the latter are those that improve the resolution only modestly (up to about a factor of two) beyond the diffraction-limit, such as confocal microscopy with closed pinhole or aided by computational methods such as deconvolution or detector-based pixel reassignment (e.g. re-scan microscopy, pixel reassignment), the 4Pi microscope, and structured-illumination microscopy technologies such as SIM and SMI. There are two major groups of methods for super-resolution microscopy in the far-field that can improve the resolution by a much larger fa ...
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Super-resolution Microscopy
Super-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, which is due to the diffraction of light. Super-resolution imaging techniques rely on the near-field (photon-tunneling microscopy as well as those that utilize the Pendry Superlens and near field scanning optical microscopy) or on the far-field. Among techniques that rely on the latter are those that improve the resolution only modestly (up to about a factor of two) beyond the diffraction-limit, such as confocal microscopy with closed pinhole or aided by computational methods such as deconvolution or detector-based pixel reassignment (e.g. re-scan microscopy, pixel reassignment), the 4Pi microscope, and structured-illumination microscopy technologies such as SIM and SMI. There are two major groups of methods for super-resolution microscopy in the far-field that can improve the resolution by a much larger fa ...
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Photoactivated Localization Microscopy
Photo-activated localization microscopy (PALM or FPALM) and stochastic optical reconstruction microscopy (STORM) are widefield (as opposed to point scanning techniques such as laser scanning confocal microscopy) fluorescence microscopy imaging methods that allow obtaining images with a resolution beyond the diffraction limit. The methods were proposed in 2006 in the wake of a general emergence of optical super-resolution microscopy methods, and were featured as Methods of the Year for 2008 by the ''Nature Methods'' journal. The development of PALM as a targeted biophysical imaging method was largely prompted by the discovery of new species and the engineering of mutants of fluorescent proteins displaying a controllable photochromism, such as photo-activatible GFP. However, the concomitant development of STORM, sharing the same fundamental principle, originally made use of paired cyanine dyes. One molecule of the pair (called activator), when excited near its absorption maximum, ser ...
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Diffraction Limit
The resolution of an optical imaging system a microscope, telescope, or camera can be limited by factors such as imperfections in the lenses or misalignment. However, there is a principal limit to the resolution of any optical system, due to the physics of diffraction. An optical system with resolution performance at the instrument's theoretical limit is said to be diffraction-limited. The diffraction-limited angular resolution of a telescopic instrument is inversely proportional to the wavelength of the light being observed, and proportional to the diameter of its objective's entrance aperture. For telescopes with circular apertures, the size of the smallest feature in an image that is diffraction limited is the size of the Airy disk. As one decreases the size of the aperture of a telescopic lens, diffraction proportionately increases. At small apertures, such as f/22, most modern lenses are limited only by diffraction and not by aberrations or other imperfections in the co ...
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Photoswitch
A photoswitch is a type of molecule that can change its structural geometry and chemical properties upon irradiation with electromagnetic radiation. Although often used interchangeably with the term molecular machine, a switch does not perform work upon a change in its shape whereas a machine does. However, photochromic compounds are the necessary building blocks for light driven molecular motors and machines. Upon irradiation with light, photoisomerization about double bonds in the molecule can lead to changes in the cis- or trans- configuration. These photochromic molecules are being considered for a range of applications. Chemical structures and properties A photochromic compound can change its configuration or structure upon irradiation with light. Several examples of photochromic compounds include: azobenzene, spiropyran, merocyanine, diarylethene, spirooxazine, fulgide, hydrazone, nobormadiene, thioindigo, acrylamide-azobenzene-quaternary ammonia, donor-acceptor Stenhouse ...
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Cumulant
In probability theory and statistics, the cumulants of a probability distribution are a set of quantities that provide an alternative to the '' moments'' of the distribution. Any two probability distributions whose moments are identical will have identical cumulants as well, and vice versa. The first cumulant is the mean, the second cumulant is the variance, and the third cumulant is the same as the third central moment. But fourth and higher-order cumulants are not equal to central moments. In some cases theoretical treatments of problems in terms of cumulants are simpler than those using moments. In particular, when two or more random variables are statistically independent, the -th-order cumulant of their sum is equal to the sum of their -th-order cumulants. As well, the third and higher-order cumulants of a normal distribution are zero, and it is the only distribution with this property. Just as for moments, where ''joint moments'' are used for collections of random variab ...
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Sofi2cc
Sofi may refer to: *Sofi (mascot), the Mascot for 2010 ISF Women's World Championship *Sofi Marinova (born 1975), Bulgarian singer SOFI may refer to: *Swedish Institute for Language and Folklore * Spray-On Foam Insulation, used on the Space Shuttle *Social finance, mobilizing investment capital to drive social progress *SoFi (Social Finance Inc.), an online personal finance company *Sofia Toufa (born 1983), also known by her stage name SOFI *South of Fifth, a neighborhood in Miami Beach, FL, also known as SOFI *SoFi Stadium, in Los Angeles, CA *Super-resolution optical fluctuation imaging Super-resolution optical fluctuation imaging (SOFI) is a post-processing method for the calculation of super-resolved images from recorded image time series that is based on the temporal correlations of independently fluctuating fluorescent emitters ...
(SOFI), a technique for super-resolution microscopy {{disambiguation ...
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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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Image Processing
An image is a visual representation of something. It can be two-dimensional, three-dimensional, or somehow otherwise feed into the visual system to convey information. An image can be an artifact, such as a photograph or other two-dimensional picture, that resembles a subject. In the context of signal processing, an image is a distributed amplitude of color(s). In optics, the term “image” may refer specifically to a 2D image. An image does not have to use the entire visual system to be a visual representation. A popular example of this is of a greyscale image, which uses the visual system's sensitivity to brightness across all wavelengths, without taking into account different colors. A black and white visual representation of something is still an image, even though it does not make full use of the visual system's capabilities. Images are typically still, but in some cases can be moving or animated. Characteristics Images may be two or three-dimensional, such as a pho ...
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