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Immunostain
In biochemistry, immunostaining is any use of an antibody-based method to detect a specific protein in a sample. The term "immunostaining" was originally used to refer to the immunohistochemical staining of tissue sections, as first described by Albert Coons in 1941. However, immunostaining now encompasses a broad range of techniques used in histology, cell biology, and molecular biology that use antibody-based staining methods. Techniques Immunohistochemistry Immunohistochemistry or IHC staining of tissue sections (or immunocytochemistry, which is the staining of cells), is perhaps the most commonly applied immunostaining technique. While the first cases of IHC staining used fluorescent dyes (see ''immunofluorescence''), other non-fluorescent methods using enzymes such as peroxidase (see '' immunoperoxidase staining'') and alkaline phosphatase are now used. These enzymes are capable of catalysing reactions that give a coloured product that is easily detectable by light micr ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Immunoperoxidase
{{Short description, Type of immunostain Immunoperoxidase is a type of immunostain used in molecular biology, medical research, and clinical diagnostics. In particular, immunoperoxidase reactions refer to a sub-class of immunohistochemical or immunocytochemical procedures in which the antibodies are visualized via a peroxidase-catalyzed reaction. Immunohistochemistry and immunocytochemistry are methods used to determine in which cells or parts of cells, a particular protein or other macromolecule are located. These stains use ''antibodies'' to bind to specific ''antigens'', usually of protein or glycoprotein origin. Since antibodies are normally invisible, special strategies must be employed to detect these bound antibodies. In an immunoperoxidase procedure, an enzyme known as a peroxidase is used to catalyze a chemical reaction to produce a coloured product. Simply, a very thin slice of tissue is fixed onto glass, incubated with antibody or a series of antibodies, the last of whic ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Immunohistochemical Staining
Immunohistochemistry is a form of immunostaining. It involves the process of selectively identifying antigens in cells and tissue, by exploiting the principle of antibodies binding specifically to antigens in biological tissues. Albert Hewett Coons, Ernest Berliner, Norman Jones and Hugh J Creech was the first to develop immunofluorescence in 1941. This led to the later development of immunohistochemistry. Immunohistochemical staining is widely used in the diagnosis of abnormal cells such as those found in cancerous tumors. In some cancer cells certain tumor antigens are expressed which make it possible to detect. Immunohistochemistry is also widely used in basic research, to understand the distribution and localization of biomarkers and differentially expressed proteins in different parts of a biological tissue. Sample preparation Immunohistochemistry can be performed on tissue that has been fixed and embedded in paraffin, but also cryopreservated (frozen) tissue. Based on ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Microscopy
Microscopy is the technical field of using microscopes to view subjects too small to 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 microscope, optical, electron microscope, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy. Optical microscopy and electron microscopy involve the diffraction, reflection (physics), reflection, or refraction of electromagnetic radiation/electron beams interacting with the Laboratory specimen, 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 microscope, transmission electron microscopy) or by scanning a fine beam over the sample (for example confocal laser scanning microscopy and scanning electron microscopy). Scan ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Immunofluorescence
Immunofluorescence (IF) is a light microscopy-based technique that allows detection and localization of a wide variety of target biomolecules within a cell or tissue at a quantitative level. The technique utilizes the binding specificity of antibodies and antigens. The specific region an antibody recognizes on an antigen is called an epitope. Several antibodies can recognize the same epitope but differ in their binding affinity. The antibody with the higher affinity for a specific epitope will surpass antibodies with a lower affinity for the same epitope. By conjugating the antibody to a fluorophore, the position of the target biomolecule is visualized by exciting the fluorophore and measuring the emission of light in a specific predefined wavelength using a fluorescence microscope. It is imperative that the binding of the fluorophore to the antibody itself does not interfere with the immunological specificity of the antibody or the binding capacity of its antigen. Immunofluore ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Chemical Synthesis
Chemical synthesis (chemical combination) is the artificial execution of chemical reactions to obtain one or several products. This occurs by physical and chemical manipulations usually involving one or more reactions. In modern laboratory uses, the process is reproducible and reliable. A chemical synthesis involves one or more compounds (known as '' reagents'' or ''reactants'') that will experience a transformation under certain conditions. Various reaction types can be applied to formulate a desired product. This requires mixing the compounds in a reaction vessel, such as a chemical reactor or a simple round-bottom flask. Many reactions require some form of processing (" work-up") or purification procedure to isolate the final product. The amount produced by chemical synthesis is known as the '' reaction yield''. Typically, yields are expressed as a mass in grams (in a laboratory setting) or as a percentage of the total theoretical quantity that could be produced based ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Gel Electrophoresis
Gel electrophoresis is an electrophoresis method for separation and analysis of biomacromolecules (DNA, RNA, proteins, etc.) and their fragments, based on their size and charge through a gel. It is used in clinical chemistry to separate proteins by charge or size (IEF agarose, essentially size independent) and in biochemistry and molecular biology to separate a mixed population of DNA and RNA fragments by length, to estimate the size of DNA and RNA fragments, or to separate proteins by charge. Nucleic acid molecules are separated by applying an electric field to move the negatively charged molecules through a gel matrix of agarose, polyacrylamide, or other substances. Shorter molecules move faster and migrate farther than longer ones because shorter molecules migrate more easily through the pores of the gel. This phenomenon is called sieving. Proteins are separated by the charge in agarose because the pores of the gel are too large to sieve proteins. Gel electrophoresi ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Protein Purification
Protein purification is a series of processes intended to isolate one or a few proteins from a complex mixture, usually Cell biology, cells, Tissue (biology), tissues, or whole organisms. Protein purification is vital for the specification of the function, structure, and interactions of the protein of interest. The purification process may separate the protein and non-protein parts of the mixture, and finally separate the desired protein from all other proteins. Ideally, to study a protein of interest, it must be separated from other components of the cell so that contaminants will not interfere in the examination of the protein of interest's structure and function. Separation of one protein from all others is typically the most laborious aspect of protein purification. Separation steps usually exploit differences in protein size, physico-chemical properties, binding affinity, and biological activity. The pure result may be termed protein isolate. Purpose The protein manufacturin ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Flow Cytometry
Flow cytometry (FC) is a technique used to detect and measure the physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument. The sample is focused to ideally flow one cell at a time through a laser beam, where the light scattered is characteristic to the cells and their components. Cells are often labeled with fluorescent markers so light is absorbed and then emitted in a band of wavelengths. Tens of thousands of cells can be quickly examined and the data gathered are processed by a computer. Flow cytometry is routinely used in basic research, clinical practice, and clinical trials. Uses for flow cytometry include: * Cell counting * Cell sorting * Determining cell characteristics and function * Detecting microorganisms * Biomarker detection * Protein engineering detection * Diagnosis of health disorders such as blood cancers * Me ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Scientific Control
A scientific control is an experiment or observation designed to minimize the effects of variables other than the independent variable (i.e. confounding variables). This increases the reliability of the results, often through a comparison between control measurements and the other measurements. Scientific controls are a part of the scientific method. Controlled experiments Controls eliminate alternate explanations of experimental results, especially experimental errors and experimenter bias. Many controls are specific to the type of experiment being performed, as in the molecular markers used in SDS-PAGE experiments, and may simply have the purpose of ensuring that the equipment is working properly. The selection and use of proper controls to ensure that experimental results are valid (for example, absence of confounding variables) can be very difficult. Control measurements may also be used for other purposes: for example, a measurement of a microphone's background noise in ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Vibratome
A microtome (from the Greek ''mikros'', meaning "small", and ''temnein'', meaning "to cut") is a cutting tool used to produce extremely thin slices of material known as ''sections'', with the process being termed microsectioning. Important in science, microtomes are used in microscopy for the preparation of samples for observation under transmitted light or electron radiation. Microtomes use steel, glass or diamond blades depending upon the specimen being sliced and the desired thickness of the sections being cut. Steel blades are used to prepare histological sections of animal or plant tissues for light microscopy. Glass knives are used to slice sections for light microscopy and to slice very thin sections for electron microscopy. Industrial grade diamond knives are used to slice hard materials such as bone, teeth and tough plant matter for both light microscopy and for electron microscopy. Gem-quality diamond knives are also used for slicing thin sections for electron microsc ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Liquid Nitrogen
Liquid nitrogen (LN2) is nitrogen in a liquid state at cryogenics, low temperature. Liquid nitrogen has a boiling point of about . It is produced industrially by fractional distillation of liquid air. It is a colorless, mobile liquid whose viscosity is about one-tenth that of acetone (i.e. roughly one-thirtieth that of water at room temperature). Liquid nitrogen is widely used as a coolant. Physical properties The diatomic character of the N2 molecule is retained after liquefaction. The weak van der Waals interaction between the N2 molecules results in little interatomic attraction. This is the cause of nitrogen's unusually low boiling point. The temperature of liquid nitrogen can readily be reduced to its freezing point by placing it in a vacuum chamber pumped by a vacuum pump. Liquid nitrogen's efficiency as a coolant is limited by the fact that it boils immediately on contact with a warmer object, enveloping the object in an insulating layer of nitrogen gas bubbles. Thi ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Paraffin Wax
Paraffin wax (or petroleum wax) is a soft colorless solid derived from petroleum, coal, or oil shale that consists of a mixture of hydrocarbon molecules containing between 20 and 40 carbon atoms. It is solid at room temperature and melting point, begins to melt above approximately , and its boiling point is above . Common applications for paraffin wax include lubrication, electrical insulation, and candles; dyed paraffin wax can be made into crayons. Un-dyed, unscented paraffin candles are odorless and bluish-white. Paraffin wax was first created by Carl Reichenbach#Scientific contributions, Carl Reichenbach in Germany in 1830 and marked a major advancement in candlemaking technology, as it burned more cleanly and reliably than tallow candles and was cheaper to produce. In chemistry, ''paraffin'' is used synonymously with ''alkane'', indicating hydrocarbons with the general formula C''n''H2''n''+2. The name is derived from Latin ''parum'' ("very little") + ''affinis'', meaning ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
