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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 However, the concomitant development of STORM, sharing the same fundamental principle, originally made use of paired cyanine dyes.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 maximu ...
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Laser Scanning 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 ...
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Normal Distribution
In statistics, a normal distribution or Gaussian distribution is a type of continuous probability distribution for a real-valued random variable. The general form of its probability density function is : f(x) = \frac e^ The parameter \mu is the mean or expectation of the distribution (and also its median and mode), while the parameter \sigma is its standard deviation. The variance of the distribution is \sigma^2. A random variable with a Gaussian distribution is said to be normally distributed, and is called a normal deviate. Normal distributions are important in statistics and are often used in the natural and social sciences to represent real-valued random variables whose distributions are not known. Their importance is partly due to the central limit theorem. It states that, under some conditions, the average of many samples (observations) of a random variable with finite mean and variance is itself a random variable—whose distribution converges to a normal d ...
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Journal
A journal, from the Old French ''journal'' (meaning "daily"), may refer to: *Bullet journal, a method of personal organization *Diary, a record of what happened over the course of a day or other period *Daybook, also known as a general journal, a daily record of financial transactions *Logbook, a record of events important to the operation of a vehicle, facility, or otherwise *Record (other) *Transaction log, a chronological record of data processing * Travel journal In publishing, ''journal'' can refer to various periodicals or serials: *Academic journal, an academic or scholarly periodical **Scientific journal, an academic journal focusing on science **Medical journal, an academic journal focusing on medicine **Law review, a professional journal focusing on legal interpretation *Magazine, non-academic or scholarly periodicals in general **Trade magazine, a magazine of interest to those of a particular profession or trade **Literary magazine, a magazine devoted to litera ...
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Pointillistic
Pointillism (, ) is a technique of painting in which small, distinct dots of color are applied in patterns to form an image. Georges Seurat and Paul Signac developed the technique in 1886, branching from Impressionism. The term "Pointillism" was coined by art critics in the late 1880s to ridicule the works of these artists, but is now used without its earlier pejorative connotation. The movement Seurat began with this technique is known as Neo-impressionism. The Divisionists used a similar technique of patterns to form images, though with larger cube-like brushstrokes. Technique The technique relies on the ability of the eye and mind of the viewer to blend the color spots into a fuller range of tones. It is related to Divisionism, a more technical variant of the method. Divisionism is concerned with color theory, whereas pointillism is more focused on the specific style of brushwork used to apply the paint. It is a technique with few serious practitioners today and is notably ...
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Light Sheet Fluorescence Microscopy
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 partic ...
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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 th ...
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Signal-to-noise Ratio
Signal-to-noise ratio (SNR or S/N) is a measure used in science and engineering that compares the level of a desired signal to the level of background noise. SNR is defined as the ratio of signal power to the noise power, often expressed in decibels. A ratio higher than 1:1 (greater than 0 dB) indicates more signal than noise. SNR, bandwidth, and channel capacity of a communication channel are connected by the Shannon–Hartley theorem. Definition Signal-to-noise ratio is defined as the ratio of the power of a signal (meaningful input) to the power of background noise (meaningless or unwanted input): : \mathrm = \frac, where is average power. Both signal and noise power must be measured at the same or equivalent points in a system, and within the same system bandwidth. Depending on whether the signal is a constant () or a random variable (), the signal-to-noise ratio for random noise becomes: : \mathrm = \frac where E refers to the expected value, i.e. in this ...
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CMOS
Complementary metal–oxide–semiconductor (CMOS, pronounced "sea-moss", ) is a type of metal–oxide–semiconductor field-effect transistor (MOSFET) fabrication process that uses complementary and symmetrical pairs of p-type and n-type MOSFETs for logic functions. CMOS technology is used for constructing integrated circuit (IC) chips, including microprocessors, microcontrollers, memory chips (including CMOS BIOS), and other digital logic circuits. CMOS technology is also used for analog circuits such as image sensors (CMOS sensors), data converters, RF circuits ( RF CMOS), and highly integrated transceivers for many types of communication. The CMOS process was originally conceived by Frank Wanlass at Fairchild Semiconductor and presented by Wanlass and Chih-Tang Sah at the International Solid-State Circuits Conference in 1963. Wanlass later filed US patent 3,356,858 for CMOS circuitry and it was granted in 1967. commercialized the technology with the trademark ...
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Charge-coupled Device
A charge-coupled device (CCD) is an integrated circuit containing an array of linked, or coupled, capacitors. Under the control of an external circuit, each capacitor can transfer its electric charge to a neighboring capacitor. CCD sensors are a major technology used in digital imaging. In a CCD image sensor, pixels are represented by p-doped metal–oxide–semiconductor (MOS) capacitors. These MOS capacitors, the basic building blocks of a CCD, are biased above the threshold for inversion when image acquisition begins, allowing the conversion of incoming photons into electron charges at the semiconductor-oxide interface; the CCD is then used to read out these charges. Although CCDs are not the only technology to allow for light detection, CCD image sensors are widely used in professional, medical, and scientific applications where high-quality image data are required. In applications with less exacting quality demands, such as consumer and professional digital cameras, a ...
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