Supravital Stain
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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 cells the r ...
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Reticulocytes Human Blood Supravital Stain
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 volume. The normal ...
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Propidium Iodide
Propidium iodide (or PI) is a fluorescent intercalating agent that can be used to stain 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 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 necrotic, apoptotic and healthy cells based on membrane integrity. PI also binds to RNA, necessitating treatment with nucleases to distinguish between RNA and DNA staining. PI is widely used in fluorescence stai ...
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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. They are all soluble in water, ethanol, diethylene glycol and dipropylene glycol. Methyl violet 2B Methyl violet 2B (IUPAC name: ''N''-(4-(bis(4-(dimethylamino)phenyl)methylene)cyclohexa-2,5-dien-1-ylidene)methanaminium chloride) is a green powder which is soluble in water and ethanol but not in xylene. It appears yellow in solution of low pH (~0.15) and changes to violet with pH increasing toward 3.2. Methyl violet 10B Methyl vi ...
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Crystal Violet
Crystal violet or gentian violet, also known as methyl violet 10B or hexamethyl pararosaniline chloride, is a triarylmethane dye used as a histological stain and in Gram's method of classifying bacteria. Crystal violet has antibacterial, antifungal, and 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 gentian flowers; it is not made from gentians or violets. Production A number of possible routes can be used to prepare crystal violet. The original procedure developed by the German chemists Kern and Caro involved the reaction of dimethylaniline with phosgene to gi ...
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Brilliant Cresyl Blue
Brilliant cresyl blue is a supravital stain used for counting reticulocytes. It is classified as an oxazine dye Oxazines are heterocyclic compounds containing one oxygen and one nitrogen atom in a doubly unsaturated six-membered ring. Isomers exist depending on the relative position of the heteroatoms and relative position of the double bonds. By extensi .... 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. Methylene blue is a thiazine dye. As a medication, it is mainly used to treat methemoglobinemia by converting the ferric iron in hemoglobin ... 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) is a non-SI unit of mass widely used in physics and chemistry. It is defined as of the mass of an unbound neutral atom of carbon-12 in its nuclear and electronic ground state and at rest. The atomic mass constant, denoted ''m''u, is defined identically, giving . This unit is commonly used in physics and chemistry to express the mass of atomic-scale objects, such as atoms, molecules, and elementary particles, both for discrete instances and multiple types of ensemble averages. For example, an atom of helium-4 has a mass of . This is an intrinsic property of the isotope and all helium-4 atoms have the same mass. Acetylsalicylic acid (aspirin), , has an average mass of approximately . However, there are no acetylsalicylic acid molecules with this mass. The two most common masses of individual acetylsalicylic acid molecules are , having the most common isotopes, and , in which one carbon is carbon-13. The molecular mass ...
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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 to be functionally interchangeable for most applications. Triton X-100 was originally a registered trademark of Rohm & 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 of a ...
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Cell Permeabilization
Cell most often refers to: * Cell (biology), the functional basic unit of life Cell may also refer to: Locations * Monastic cell, a small room, hut, or cave in which a religious recluse lives, alternatively the small precursor of a monastery with only a few monks or nuns * Prison cell, a room used to hold people in prisons Groups of people * Cell, a group of people in a cell group, a form of Christian church organization * Cell, a unit of a clandestine cell system, a penetration-resistant form of a secret or outlawed organization * Cellular organizational structure, such as in business management Science, mathematics, and technology Computing and telecommunications * Cell (EDA), a term used in an electronic circuit design schematics * Cell (microprocessor), a microprocessor architecture developed by Sony, Toshiba, and IBM * Memory cell (computing), the basic unit of (volatile or non-volatile) computer memory * Cell, a unit in a database table or spreadsheet, formed by the ...
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Fluorescence-activated Cell Sorting
Flow cytometry (FC) is a technique used to detect and measure 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 * Measuring ...
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Immunofluorescence
Immunofluorescence is a technique used for light microscopy with a fluorescence microscope and is used primarily on microbiological samples. This technique uses the specificity of antibodies to their antigen to target fluorescent dyes to specific biomolecule targets within a cell, and therefore allows visualization of the distribution of the target molecule through the sample. The specific region an antibody recognizes on an antigen is called an epitope. There have been efforts in epitope mapping since many antibodies can bind the same epitope and levels of binding between antibodies that recognize the same epitope can vary. Additionally, the binding of the fluorophore to the antibody itself cannot interfere with the immunological specificity of the antibody or the binding capacity of its antigen. Immunofluorescence is a widely used example of immunostaining (using antibodies to stain proteins) and is a specific example of immunohistochemistry (the use of the antibody-antigen rel ...
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