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Miles And Misra Method
The Miles and Misra Method (or surface viable count) is a technique used in Microbiology to determine the number of colony forming units in a bacterial suspension or homogenate. The technique was first described in 1938 by Miles, Misra and Irwin who at the time were working at the LSHTM. The Miles and Misra method has been shown to be precise. Materials * A calibrated dropping pipette, or automatic pipette, delivering drops of 20μl. * Petri dishes containing nutrient agar or other appropriate medium. * Phosphate Buffered Saline (PBS) or other appropriate diluent. * Bacterial suspension or homogenate. Method *The inoculum / suspension is serially diluted by adding 1x of suspension to 9x of diluent. When the quantity of bacteria is unknown, dilutions should be made to at least 10−8. *Three plates are needed for each dilution series, for statistical reasons an average of at least 3 counts are needed. *The surface of the plates need to be sufficiently dry to allow a 20μl drop to ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Microbiology
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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Colony Forming Unit
In microbiology, colony-forming unit (CFU, cfu or Cfu) is a unit which estimates the number of microbial cells (bacteria, fungi, viruses etc.) in a sample that are viable, able to multiply via binary fission under the controlled conditions. Counting with colony-forming units requires culturing the microbes and counts only viable cells, in contrast with microscopic examination which counts all cells, living or dead. The visual appearance of a colony in a cell culture requires significant growth, and when counting colonies, it is uncertain if the colony arose from one cell or a group of cells. Expressing results as colony-forming units reflects this uncertainty. Theory The purpose of plate counting is to estimate the number of cells present based on their ability to give rise to colonies under specific conditions of nutrient medium, temperature and time. Theoretically, one viable cell can give rise to a colony through replication. However, solitary cells are the exception in natur ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Bacterial
Bacteria (; singular: bacterium) are ubiquitous, mostly free-living organisms often consisting of one biological cell. They constitute a large domain of prokaryotic microorganisms. Typically a few micrometres in length, bacteria were among the first life forms to appear on Earth, and are present in most of its habitats. Bacteria inhabit soil, water, acidic hot springs, radioactive waste, and the deep biosphere of Earth's crust. Bacteria are vital in many stages of the nutrient cycle by recycling nutrients such as the fixation of nitrogen from the atmosphere. The nutrient cycle includes the decomposition of dead bodies; bacteria are responsible for the putrefaction stage in this process. In the biological communities surrounding hydrothermal vents and cold seeps, extremophile bacteria provide the nutrients needed to sustain life by converting dissolved compounds, such as hydrogen sulphide and methane, to energy. Bacteria also live in symbiotic and parasitic relationships ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Homogenization (biology)
Homogenization, in cell biology or molecular biology, is a process whereby different fractions of a biological sample become equal in composition. It can be a disease sign in histopathology, or an intentional process in research: A homogenized sample is equal in composition throughout, so that removing a fraction does not alter the overall molecular make-up of the sample remaining, and is identical to the fraction removed. Induced homogenization in biology is often followed by molecular extraction and various analytical techniques, including ELISA and western blot. Methods Homogenization of tissue in solution is often performed simultaneously with cell lysis. To prevent lysis however, the tissue (or collection of cells, e.g. from cell culture) can be kept at temperatures slightly above zero to prevent autolysis, and in an isotonic solution to prevent osmotic damage. If freezing the tissue is possible, cryohomogenization can be performed under "dry" conditions, and is often the m ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
London School Of Hygiene & Tropical Medicine
The London School of Hygiene and Tropical Medicine (LSHTM) is a public research university in Bloomsbury, central London, and a member institution of the University of London that specialises in public health and tropical medicine. The institution was founded in 1899 by Sir Patrick Manson, after a donation from the Indian Parsi philanthropist B. D. Petit. Since its foundation it has become one of the most highly placed institutions in global rankings in the fields of public health and infectious diseases. The annual income of the institution for 2020–21 was £244.2 million, of which £167.6 million was from research grants and contracts, with expenditures totalling £235.2 million during the same period. History Origins (1899–1913) The school was founded on October 2, 1899, by Sir Patrick Manson as the London School of Tropical Medicine after the Parsi philanthropist Bomanjee Dinshaw Petit made a donation of £6,666. It was initially located at ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Pipette
A pipette (sometimes spelled as pipett) is a laboratory tool commonly used in chemistry, biology and medicine to transport a measured volume of liquid, often as a media dispenser. Pipettes come in several designs for various purposes with differing levels of accuracy and precision, from single piece glass pipettes to more complex adjustable or electronic pipettes. Many pipette types work by creating a partial vacuum above the liquid-holding chamber and selectively releasing this vacuum to draw up and dispense liquid. Measurement accuracy varies greatly depending on the instrument. History The first simple pipettes were made in glass, such as Pasteur pipettes. Large pipettes continue to be made in glass; others are made in squeezable plastic for situations where an exact volume is not required. The first micropipette was patented in 1957 by Dr Heinrich Schnitger (Marburg, Germany). The founder of the company Eppendorf, Dr. Heinrich Netheler, inherited the rights and started the ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Petri Dish
A Petri dish (alternatively known as a Petri plate or cell-culture dish) is a shallow transparent lidded dish that biologists use to hold growth medium in which cells can be cultured,R. C. Dubey (2014): ''A Textbook Of Biotechnology For Class-XI'', 4th edition, p. 469. originally, cells of bacteria, fungi and small mosses. The container is named after its inventor, German bacteriologist Julius Richard Petri. It is the most common type of culture plate. The Petri dish is one of the most common items in biology laboratories and has entered popular culture. The term is sometimes written in lower case, especially in non-technical literature. What was later called Petri dish was originally developed by German physician Robert Koch in his private laboratory in 1881, as a precursor method. Petri, as assistant to Koch, at Berlin University made the final modifications in 1887 as used today. Penicillin, the first antibiotic, was discovered in 1929 when Alexander Fleming noticed that m ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Agar
Agar ( or ), or agar-agar, is a jelly-like substance consisting of polysaccharides obtained from the cell walls of some species of red algae, primarily from ogonori (''Gracilaria'') and "tengusa" (''Gelidiaceae''). As found in nature, agar is a mixture of two components, the linear polysaccharide agarose and a heterogeneous mixture of smaller molecules called agaropectin. It forms the supporting structure in the cell walls of certain species of algae and is released on boiling. These algae are known as agarophytes, belonging to the Rhodophyta (red algae) phylum. The processing of food-grade agar removes the agaropectin, and the commercial product is essentially pure agarose. Agar has been used as an ingredient in desserts throughout Asia and also as a solid substrate to contain culture media for microbiological work. Agar can be used as a laxative; an appetite suppressant; a vegan substitute for gelatin; a thickener for soups; in fruit preserves, ice cream, and other desser ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Phosphate Buffered Saline
Phosphate-buffered saline (PBS) is a buffer solution (pH ~ 7.4) commonly used in biological research. It is a water-based salt solution containing disodium hydrogen phosphate, sodium chloride and, in some formulations, potassium chloride and potassium dihydrogen phosphate. The buffer helps to maintain a constant pH. The osmolarity and ion concentrations of the solutions match those of the human body ( isotonic). Applications PBS has many uses because it is isotonic and non-toxic to most cells. These uses include substance dilution and cell container rinsing. PBS with EDTA is also used to disengage attached and clumped cells. Divalent metals such as zinc, however, cannot be added as this will result in precipitation. For these types of applications, Good's buffers are recommended. PBS has been shown to be an acceptable alternative to viral transport medium regarding transport and storage of RNA viruses, such as SARS-CoV-2 Preparation There are many different ways to prepare PBS so ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Colony-forming Unit
In microbiology, colony-forming unit (CFU, cfu or Cfu) is a unit which estimates the number of microbial cells (bacteria, fungi, viruses etc.) in a sample that are viable, able to multiply via binary fission under the controlled conditions. Counting with colony-forming units requires culturing the microbes and counts only viable cells, in contrast with microscopic examination which counts all cells, living or dead. The visual appearance of a colony in a cell culture requires significant growth, and when counting colonies, it is uncertain if the colony arose from one cell or a group of cells. Expressing results as colony-forming units reflects this uncertainty. Theory The purpose of plate counting is to estimate the number of cells present based on their ability to give rise to colonies under specific conditions of nutrient medium, temperature and time. Theoretically, one viable cell can give rise to a colony through replication. However, solitary cells are the exception in na ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Cell Culture Techniques
Cell culture or tissue culture is the process by which cells are grown under controlled conditions, generally outside of their natural environment. The term "tissue culture" was coined by American pathologist Montrose Thomas Burrows. This technique is also called micropropagation. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions the need to be kept at body temperature (37 °C) in an incubator. These conditions vary for each cell type, but generally consist of a suitable vessel with a substrate or rich medium that supplies the essential nutrients (amino acids, carbohydrates, vitamins, minerals), growth factors, hormones, and gases ( CO2, O2), and regulates the physio-chemical environment (pH buffer, osmotic pressure, temperature). Most cells require a surface or an artificial substrate to form an adherent culture as a monolayer (one single-cell thick), whereas others can be grown ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
