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Supravital Stain
Supravital staining is a method of staining used in microscopy to examine living cells that have been removed from an organism. It differs from intravital staining, which is done by injecting or otherwise introducing the stain into the body. Thus a supravital stain may have a greater toxicity, as only a few cells need to survive it a short while. The term " vital stain" is used by some authors to refer specifically to an intravital stain, and by others interchangeably with a supravital stain, the core concept being that the cell being examined is still alive. As the cells are alive and unfixed, outside the body, supravital stains are temporary in nature. The most common supravital stain is performed on reticulocytes using new methylene blue or brilliant cresyl blue, which makes it possible to see the reticulofilamentous pattern of ribosomes characteristically precipitated in these live immature red blood cells by the supravital stains. By counting the number of such cell ...
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Reticulocytes Human Blood Supravital Stain
In hematology, reticulocytes are immature red blood cells (RBCs). In the process of erythropoiesis (red blood cell formation), reticulocytes develop and mature in the bone marrow and then circulate for about a day in the blood stream before developing into mature red blood cells. Like mature red blood cells, in mammals, reticulocytes do not have a cell nucleus. They are called reticulocytes because of a reticular (mesh-like) network of ribosomal RNA that becomes visible under a microscope with certain stains such as new methylene blue and Romanowsky stain. Clinical significance To accurately measure reticulocyte counts, automated counters use a combination of laser excitation, detectors and a fluorescent dye that marks RNA and DNA (such as titan yellow or polymethine). Reticulocytes appear slightly bluer than other red cells when looked at with the normal Romanowsky stain. Reticulocytes are also relatively large, a characteristic that is described by the mean corpuscular v ...
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Propidium Iodide
Propidium iodide (or PI) is a Fluorescence#Biochemistry and medicine, fluorescent intercalating agent that can be used to Staining (biology), stain cell (biology), cells and nucleic acids. PI binds to DNA by intercalating between the bases with little or no sequence preference. When in an aqueous solution, PI has a fluorescent excitation maximum of 493 nm (blue-green), and an emission maximum of 636 nm (red). After binding DNA, the quantum yield of PI is enhanced 20-30 fold, and the excitation/emission maximum of PI is shifted to 535 nm (green) / 617 nm (orange-red). Propidium iodide is used as a DNA stain in flow cytometry to Viability assay, evaluate cell viability or DNA content in cell cycle analysis, or in microscopy to visualize the nucleus and other DNA-containing organelles. Propidium Iodide is not membrane-permeable, making it useful to differentiate Necrosis, necrotic, Apoptosis, apoptotic and healthy cells based on membrane integrity. PI also binds to RNA, necessitating ...
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Methyl Violet
Methyl violet is a family of organic compounds that are mainly used as dyes. Depending on the number of attached methyl groups, the color of the dye can be altered. Its main use is as a purple dye for textiles and to give deep violet colors in paint and ink. It is also used as a hydration indicator for silica gel. Methyl violet 10B is also known as crystal violet (and many other names) and has medical uses. Structure The term methyl violet encompasses three compounds that differ in the number of methyl groups attached to the amine functional group. Methyl violets are mixtures of tetramethyl (2B), pentamethyl (6B) and hexamethyl (10B) pararosanilins. They are all soluble in water, ethanol, diethylene glycol and dipropylene glycol. : Methyl violet 2B Methyl violet 2B (IUPAC name: 4,4′-((4-Iminocyclohexa-2,5-dien-1-ylidene)methylene)bis(''N'',''N''-dimethylaniline) monohydrochloride) is a green powder which is soluble in water and ethanol but not in xylene. It appears yellow in s ...
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Crystal Violet
Crystal violet or gentian violet, also known as methyl violet 10B or hexamethyl pararosaniline chloride, is a triphenylmethane, triarylmethane dye used as a histological stain and in Gram staining, Gram's method of classifying bacteria. Crystal violet has antibacterial, Antifungal medication, antifungal, and anthelmintic (Anthelmintic, vermicide) properties and was formerly important as a topical antiseptic. The medical use of the dye has been largely superseded by more modern drugs, although it is still listed by the World Health Organization. The name ''gentian violet'' was originally used for a mixture of methyl pararosaniline dyes (methyl violet), but is now often considered a synonym for ''crystal violet''. The name refers to its colour, being like that of the petals of certain Gentiana, gentian flowers; it is not made from gentians or list of plants known as violet, violets. Production A number of possible routes can be used to prepare crystal violet. The original procedur ...
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Brilliant Cresyl Blue
Brilliant cresyl blue is a supravital stain used for counting reticulocyte In hematology, reticulocytes are immature red blood cells (RBCs). In the process of erythropoiesis (red blood cell formation), reticulocytes develop and mature in the bone marrow and then circulate for about a day in the blood stream before dev ...s. It is classified as an oxazine dye. N95 dust masks, eye shields, and gloves must all be worn when handling the chemical. References {{heterocyclic-stub Chlorides Zinc compounds Oxazine dyes Phenoxazines ...
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New Methylene Blue
is an organic compound of the thiazine class of heterocycles. It is used as a stain and as an antimicrobial agent. It is classified as an azine dye, and the chromophore is a cation, the anion is often unspecified. Applications NMB is a staining agent used in diagnostic cytopathology and histopathology, typically for staining immature red blood cells. It is a supravital stain. It is closely related to methylene blue, an older stain in wide use. Safety New methylene blue is toxic. Skin contact or inhalation should be avoided. See also * Methylene blue Methylthioninium chloride, commonly called methylene blue, is a salt used as a dye and as a medication. As a medication, it is mainly used to treat methemoglobinemia. It has previously been used for treating cyanide poisoning and urinary trac ... References {{DEFAULTSORT:New Methylene Blue Biochemistry detection methods Thiazine dyes Vital stains Histology ...
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Kilodalton
The dalton or unified atomic mass unit (symbols: Da or u, respectively) is a unit of mass defined as of the mass of an unbound neutral atom of carbon-12 in its nuclear and electronic ground state and at rest. It is a non-SI unit accepted for use with SI. The word "unified" emphasizes that the definition was accepted by both IUPAP and IUPAC. The atomic mass constant, denoted , is defined identically. Expressed in terms of , the atomic mass of carbon-12: . Its value in SI units is an experimentally determined quantity. The 2022 CODATA recommended value of the atomic mass constant expressed in the SI base unit kilogram is:This value serves as a conversion factor of mass from daltons to kilograms, which can easily be converted to grams and other metric units of mass. The 2019 revision of the SI redefined the kilogram by fixing the value of the Planck constant (), improving the precision of the atomic mass constant expressed in SI units by anchoring it to fixed physical ...
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Triton X-100
Triton X-100 (''n'') is a nonionic surfactant that has a hydrophilic polyethylene oxide chain (on average it has 9.5 ethylene oxide units) and an aromatic hydrocarbon lipophilic or hydrophobic group. The hydrocarbon group is a 4-( 1,1,3,3-tetramethylbutyl)- phenyl group. Triton X-100 is closely related to IGEPAL CA-630, which might differ from it mainly in having slightly shorter ethylene oxide chains. As a result, Triton X-100 is slightly more hydrophilic than Igepal CA-630 thus these two detergents may not be considered functionally interchangeable for most applications. Triton X-100 was originally a registered trademark of Rohm and Haas Co. It was subsequently purchased by Union Carbide and then acquired by Dow Chemical Company upon the acquisition of Union Carbide. Soon afterward (in 2009), Dow also acquired Rohm & Haas Co. Physical properties Undiluted Triton X-100 is a clear viscous fluid (less viscous than undiluted glycerol). Undiluted Triton X-100 has a viscosity ...
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Cell Permeabilization
Cell most often refers to: * Cell (biology), the functional basic unit of life * Cellphone, a phone connected to a cellular network * Clandestine cell, a penetration-resistant form of a secret or outlawed organization * Electrochemical cell, a device used to convert chemical energy to electrical energy * Prison cell, a room used to hold people in prisons Cell may also refer to: Arts, entertainment, and media Fictional entities * Cell (comics), a Marvel comic book character * Cell (''Dragon Ball''), a character in the manga series ''Dragon Ball'' Literature * ''Cell'' (novel), a 2006 horror novel by Stephen King * "Cells", poem, about a hungover soldier in gaol, by Rudyard Kipling * ''The Cell'' (play), an Australian play by Robert Wales Music * Cell (music), a small rhythmic and melodic design that can be isolated, or can make up one part of a thematic context * Cell (American band) * Cell (Japanese band) * ''Cell'' (album), a 2004 album by Plastic Tree * ''Cells'', a 1998 ...
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Fluorescence-activated Cell Sorting
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 * Measu ...
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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 ...
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