Moeller Stain
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Moeller Stain
Moeller staining involves the use of a steamed dye reagent in order to increase the stainability of endospores. Carbol fuchsin is the primary stain used in this method. Endospores are stained red, while the counterstain methylene blue stains the vegetative bacteria blue. Endospores are surrounded by a highly resistant spore coat, which is highly resistant to excessive heat, freezing, desiccation, as well as chemical agents. More importantly, for identification, spores are resistant to commonly employed staining techniques; therefore alternative staining methods are required. Method Carbol fuchsin is applied to a heat-fixed slide. The slide is then heated over a bunsen burner, or suspended over a hot water bath, covered with a paper towel, and steamed for 3 minutes. The slide is rinsed with acidified ethanol, and counter-stained with Methylene blue. An improved method involves the addition of the surfactant Tergitol 7 to the carbol fuchsin stain, and the omission of the steamin ...
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Reagent
In chemistry, a reagent ( ) or analytical reagent is a substance or compound added to a system to cause a chemical reaction, or test if one occurs. The terms ''reactant'' and ''reagent'' are often used interchangeably, but reactant specifies a substance ''consumed'' in the course of a chemical reaction. ''Solvents'', though involved in the reaction mechanism, are usually not called reactants. Similarly, ''catalysts'' are not consumed by the reaction, so they are not reactants. In biochemistry, especially in connection with enzyme-catalyzed reactions, the reactants are commonly called substrates. Definitions Organic chemistry In organic chemistry, the term "reagent" denotes a chemical ingredient (a compound or mixture, typically of inorganic or small organic molecules) introduced to cause the desired transformation of an organic substance. Examples include the Collins reagent, Fenton's reagent, and Grignard reagents. Analytical chemistry In analytical chemistry, a reagent ...
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Endospores
An endospore is a dormant, tough, and non-reproductive structure produced by some bacteria in the phylum Bacillota. The name "endospore" is suggestive of a spore or seed-like form (''endo'' means 'within'), but it is not a true spore (i.e., not an offspring). It is a stripped-down, dormant form to which the bacterium can reduce itself. Endospore formation is usually triggered by a lack of nutrients, and usually occurs in gram-positive bacteria. In endospore formation, the bacterium divides within its cell wall, and one side then engulfs the other. Endospores enable bacteria to lie dormant for extended periods, even centuries. There are many reports of spores remaining viable over 10,000 years, and revival of spores millions of years old has been claimed. There is one report of viable spores of ''Bacillus marismortui'' in salt crystals approximately 250 million years old. When the environment becomes more favorable, the endospore can reactivate itself into a vegetative state. Mos ...
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Carbol Fuchsin
Carbol fuchsin, carbol-fuchsin, carbolfuchsin, or Castellani's paint ( CAS ) is a mixture of phenol and basic fuchsin that is used in bacterial staining procedures. It is commonly used in the staining of mycobacteria because it has an affinity for the mycolic acids found in their cell membranes. It is a component of Ziehl–Neelsen stain, a differential stain. Carbol fuchsin is used as the primary stain dye to detect acid-fast bacteria because it is more soluble in the cells' wall lipids than in the acid alcohol. If the bacteria is acid-fast the bacteria will retain the initial red color of the dye because they are able to resist the destaining by acid alcohol (0.4–1% HCl in 70% EtOH). Additionally, it can be used for the staining of bacterial spores. Carbol-fuchsin is also used as a topical antiseptic An antiseptic (from Greek ἀντί ''anti'', "against" and σηπτικός ''sēptikos'', "putrefactive") is an antimicrobial substance or compound that is applied to livi ...
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Counterstain
A counterstain is a stain with colour contrasting to the principal stain, making the stained structure easily visible using a microscope. Examples include the malachite green counterstain to the fuchsine stain in the Gimenez staining technique and the eosin counterstain to haematoxylin in the H&E stain. In Gram staining, crystal violet stains only Gram-positive bacteria, and safranin counterstain is applied which stains all cells, allowing the identification of Gram-negative bacteria as well. An alternative method uses dilute carbofluozide. Counterstains are sometimes used to separate animals from organic detritus In biology, detritus () is dead particulate organic material, as distinguished from dissolved organic material. Detritus typically includes the bodies or fragments of bodies of dead organisms, and fecal material. Detritus typically hosts commun ... in microbiology studies. References Staining {{pathology-stub ...
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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 to ferrous iron. Specifically, it is used to treat methemoglobin levels that are greater than 30% or in which there are symptoms despite oxygen therapy. It has previously been used for treating cyanide poisoning and urinary tract infections, but this use is no longer recommended. Methylene blue is typically given by injection into a vein. Common side effects include headache, vomiting, confusion, shortness of breath, and high blood pressure. Other side effects include serotonin syndrome, red blood cell breakdown, and allergic reactions. Use often turns the urine, sweat, and stool blue to green in color. While use during pregnancy may harm the baby, not using it in methemoglobinemia is likely more dangerous. Methylene blue was first pr ...
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Desiccation
Desiccation () is the state of extreme dryness, or the process of extreme drying. A desiccant is a hygroscopic (attracts and holds water) substance that induces or sustains such a state in its local vicinity in a moderately sealed container. Industry Desiccation is widely employed in the oil and gas industry. These materials are obtained in a hydrated state, but the water content leads to corrosion or is incompatible with downstream processing. Removal of water is achieved by cryogenic condensation, absorption into glycols, and absorption onto desiccants such as silica gel. Laboratory A desiccator is a heavy glass or plastic container, now somewhat antiquated, used in practical chemistry for drying or keeping small amounts of materials very dry. The material is placed on a shelf, and a drying agent or ''desiccant'', such as dry silica gel or anhydrous sodium hydroxide, is placed below the shelf. Often some sort of humidity indicator is included in the desiccator to show, ...
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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 to ferrous iron. Specifically, it is used to treat methemoglobin levels that are greater than 30% or in which there are symptoms despite oxygen therapy. It has previously been used for treating cyanide poisoning and urinary tract infections, but this use is no longer recommended. Methylene blue is typically given by injection into a vein. Common side effects include headache, vomiting, confusion, shortness of breath, and high blood pressure. Other side effects include serotonin syndrome, red blood cell breakdown, and allergic reactions. Use often turns the urine, sweat, and stool blue to green in color. While use during pregnancy may harm the baby, not using it in methemoglobinemia is likely more dangerous. Methylene blue was first pr ...
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Schaeffer–Fulton Stain
The Schaeffer–Fulton stain is a technique designed to isolate endospores by staining any present endospores green, and any other bacterial bodies red. The primary stain is malachite green, and the counterstain is safranin, which dyes any other bacterial bodies red. Endospores cannot be stained by normal staining procedures because their walls are practically impermeable to all chemicals. The Schaeffer- Fulton endospore stain uses heat to drive the primary stain(malachite green) into the endospore. After cooling, the slide is decolorized with water and counterstained with safranin. Procedure Using an aseptic technique, bacteria are placed on a slide and heat fixed. The slide is then suspended over a water bath with some sort of porus paper over it, so that the slide is steamed. Malachite green is applied to the slide, which can penetrate the tough walls of the endospores, staining them green. After five minutes, the slide is removed from the steam, and the paper towel is removed. ...
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Microbiology Techniques
Microbiology () is the scientific study of microorganisms, those being unicellular (single cell), multicellular (cell colony), or acellular (lacking cells). Microbiology encompasses numerous sub-disciplines including virology, bacteriology, protistology, mycology, immunology, and parasitology. Eukaryotic microorganisms possess membrane-bound organelles and include fungi and protists, whereas prokaryotic organisms—all of which are microorganisms—are conventionally classified as lacking membrane-bound organelles and include Bacteria and Archaea. Microbiologists traditionally relied on culture, staining, and microscopy. However, less than 1% of the microorganisms present in common environments can be cultured in isolation using current means. Microbiologists often rely on molecular biology tools such as DNA sequence based identification, for example the 16S rRNA gene sequence used for bacteria identification. Viruses have been variably classified as organisms, as they have bee ...
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Staining
Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Stains may be used to define biological tissues (highlighting, for example, muscle fibers or connective tissue), cell populations (classifying different blood cells), or organelles within individual cells. In biochemistry, it involves adding a class-specific ( DNA, proteins, lipids, carbohydrates) dye to a substrate to qualify or quantify the presence of a specific compound. Staining and fluorescent tagging can serve similar purposes. Biological staining is also used to mark cells in flow cytometry, and to flag proteins or nucleic acids in gel electrophoresis. Light microscopes are used for viewin ...
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Bacteriology
Bacteriology is the branch and specialty of biology that studies the morphology, ecology, genetics and biochemistry of bacteria as well as many other aspects related to them. This subdivision of microbiology involves the identification, classification, and characterization of bacterial species. Because of the similarity of thinking and working with microorganisms other than bacteria, such as protozoa, fungi, and viruses, there has been a tendency for the field of bacteriology to extend as microbiology. The terms were formerly often used interchangeably. However, bacteriology can be classified as a distinct science. Overview Definition Bacteriology is the study of bacteria and their relation to medicine. Bacteriology evolved from physicians needing to apply the germ theory to address the concerns relating to disease spreading in hospitals the 19th century. Identification and characterizing of bacteria being associated to diseases led to advances in pathogenic bacteriology. K ...
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