Nanovid Microscopy
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Nanovid Microscopy
{{Short description, Microscopic technique using colloidal gold Nanovid microscopy, from "nanometer video-enhanced microscopy", is a microscopic technique aimed at visualizing colloidal gold particles of 20–40 nm diameter (nanogold, immunogold) as dynamic markers at the light-microscopic level. The nanogold particles as such are smaller than the diffraction limit of light, but can be visualized by using video-enhanced differential interference contrast (VEDIC). The technique is based on the use of contrast enhancement by video techniques and digital image processing. Nanovid microscopy, by combining small colloidal gold probes with video-enhanced quantitative microscopy, allows studying the intracellular dynamics of specific proteins in living cells. See also * Microscopy * Single-particle tracking * Differential interference contrast microscopy * Microtubule Microtubules are polymers of tubulin that form part of the cytoskeleton and provide structure and shape to eukar ...
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Colloidal Gold
Colloidal gold is a sol or colloidal suspension of nanoparticles of gold in a fluid, usually water. The colloid is usually either wine-red coloured (for spherical particles less than 100  nm) or blue/purple (for larger spherical particles or nanorods). Due to their optical, electronic, and molecular-recognition properties, gold nanoparticles are the subject of substantial research, with many potential or promised applications in a wide variety of areas, including electron microscopy, electronics, nanotechnology, materials science, and biomedicine. The properties of colloidal gold nanoparticles, and thus their potential applications, depend strongly upon their size and shape. For example, rodlike particles have both a transverse and longitudinal absorption peak, and anisotropy of the shape affects their self-assembly. History Used since ancient times as a method of staining glass colloidal gold was used in the 4th-century Lycurgus Cup, which changes color dependin ...
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Immunogold Labelling
Immunogold labeling or Immunogold staining (IGS) is a staining technique used in electron microscopy. This staining technique is an equivalent of the indirect immunofluorescence technique for visible light. Colloidal gold particles are most often attached to secondary antibodies which are in turn attached to primary antibodies designed to bind a specific antigen or other cell component. Gold is used for its high electron density which increases electron scatter to give high contrast 'dark spots'. First used in 1971, immunogold labeling has been applied to both transmission electron microscopy and scanning electron microscopy, as well as brightfield microscopy. The labeling technique can be adapted to distinguish multiple objects by using differently-sized gold particles. Immunogold labeling can introduce artifacts, as the gold particles reside some distance from the labelled object and very thin sectioning is required during sample preparation. History Immunogold labeling was ...
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Diffraction
Diffraction is defined as the interference or bending of waves around the corners of an obstacle or through an aperture into the region of geometrical shadow of the obstacle/aperture. The diffracting object or aperture effectively becomes a secondary source of the propagating wave. Italian scientist Francesco Maria Grimaldi coined the word ''diffraction'' and was the first to record accurate observations of the phenomenon in 1660. In classical physics, the diffraction phenomenon is described by the Huygens–Fresnel principle that treats each point in a propagating wavefront as a collection of individual spherical wavelets. The characteristic bending pattern is most pronounced when a wave from a coherent source (such as a laser) encounters a slit/aperture that is comparable in size to its wavelength, as shown in the inserted image. This is due to the addition, or interference, of different points on the wavefront (or, equivalently, each wavelet) that travel by paths of d ...
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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 par ...
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Contrast (vision)
Contrast is the contradiction in luminance or colour that makes an object (or its representation in an image or display) distinguishable. In visual perception of the real world, contrast is determined by the difference in the colour and brightness of the object and other objects within the same field of view. The human visual system is more sensitive to contrast than absolute luminance; we can perceive the world similarly regardless of the huge changes in illumination over the day or from place to place. The maximum ''contrast'' of an image is the contrast ratio or dynamic range. Images with a contrast ratio close to their medium's maximum possible contrast ratio experience a ''conservation of contrast'', wherein any increase in contrast in some parts of the image must necessarily result in a decrease in contrast elsewhere. Brightening an image will increase contrast in dark areas but decrease contrast in bright areas, while darkening the image will have the opposite effect. B ...
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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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Protein
Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residues. Proteins perform a vast array of functions within organisms, including catalysing metabolic reactions, DNA replication, responding to stimuli, providing structure to cells and organisms, and transporting molecules from one location to another. Proteins differ from one another primarily in their sequence of amino acids, which is dictated by the nucleotide sequence of their genes, and which usually results in protein folding into a specific 3D structure that determines its activity. A linear chain of amino acid residues is called a polypeptide. A protein contains at least one long polypeptide. Short polypeptides, containing less than 20–30 residues, are rarely considered to be proteins and are commonly called peptides. The individual amino acid residues are bonded together by peptide bonds and adjacent amino acid residues. The sequence of amino acid residue ...
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Cell (biology)
The cell is the basic structural and functional unit of life forms. Every cell consists of a cytoplasm enclosed within a membrane, and contains many biomolecules such as proteins, DNA and RNA, as well as many small molecules of nutrients and metabolites.Cell Movements and the Shaping of the Vertebrate Body
in Chapter 21 of
Molecular Biology of the Cell
'' fourth edition, edited by Bruce Alberts (2002) published by Garland Science. The Alberts text discusses how the "cellular building blocks" move to shape developing embryos. It is also common to describe small molecules such as ...
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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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Single-particle Tracking
Single-particle tracking (SPT) is the observation of the motion of individual particles within a medium. The coordinates time series, which can be either in two dimensions (''x'', ''y'') or in three dimensions (''x'', ''y'', ''z''), is referred to as a ''trajectory''. The trajectory is typically analyzed using statistical methods to extract information about the underlying dynamics of the particle. These dynamics can reveal information about the type of transport being observed (e.g., thermal or active), the medium where the particle is moving, and interactions with other particles. In the case of random motion, trajectory analysis can be used to measure the diffusion coefficient. Applications In life sciences, single-particle tracking is broadly used to quantify the dynamics of molecules/proteins in live cells (of bacteria, yeast, mammalian cells and live ''Drosophila'' embryos). It has been extensively used to study the transcription factor dynamics in live cells. This method h ...
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Differential Interference Contrast Microscopy
Differential interference contrast (DIC) microscopy, also known as Nomarski interference contrast (NIC) or Nomarski microscopy, is an optical microscopy technique used to enhance the contrast in unstained, transparent samples. DIC works on the principle of interferometry to gain information about the optical path length of the sample, to see otherwise invisible features. A relatively complex optical system produces an image with the object appearing black to white on a grey background. This image is similar to that obtained by phase contrast microscopy but without the bright diffraction halo. The technique was developed by Polish physicist Georges Nomarski in 1952. DIC works by separating a polarized light source into two orthogonally polarized mutually coherent parts which are spatially displaced (sheared) at the sample plane, and recombined before observation. The interference of the two parts at recombination is sensitive to their optical path difference (i.e. the product ...
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Microtubule
Microtubules are polymers of tubulin that form part of the cytoskeleton and provide structure and shape to eukaryotic cells. Microtubules can be as long as 50 micrometres, as wide as 23 to 27  nm and have an inner diameter between 11 and 15 nm. They are formed by the polymerization of a dimer of two globular proteins, alpha and beta tubulin into protofilaments that can then associate laterally to form a hollow tube, the microtubule. The most common form of a microtubule consists of 13 protofilaments in the tubular arrangement. Microtubules play an important role in a number of cellular processes. They are involved in maintaining the structure of the cell and, together with microfilaments and intermediate filaments, they form the cytoskeleton. They also make up the internal structure of cilia and flagella. They provide platforms for intracellular transport and are involved in a variety of cellular processes, including the movement of secretory vesicles, organell ...
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