Percoll
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Percoll
Percoll is a tool for efficient density separation in Cell biology that was first formulated by Pertoft and colleagues. It is used for the isolation of cells, organelles, and/or viruses by density centrifugation. Percoll consists of colloidal silica particles of 15–30 nm diameter (23% w/w in water) which have been coated with polyvinylpyrrolidone (PVP). Percoll is well suited for density gradient experiments because it possesses a low viscosity compared to alternatives, a low osmolarity, and no toxicity towards cells and their constituents. Percoll is a registered trademark of GE Healthcare. Past use in artificial reproduction Percoll was previously used in assisted reproductive technology (ART) to select sperm from semen by density gradient centrifugation, for use in techniques such as ''in vitro'' fertilization or intrauterine insemination. However, in 1996, Pharmacia sent out a letter to laboratories stating that Percoll should be used for research purposes only, no ...
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Centrifugation
Centrifugation is a mechanical process which involves the use of the centrifugal force to separate particles from a solution according to their size, shape, density, medium viscosity and rotor speed. The denser components of the mixture migrate away from the axis of the centrifuge, while the less dense components of the mixture migrate towards the axis. Chemists and biologists may increase the effective gravitational force of the test tube so that the precipitate (pellet) will travel quickly and fully to the bottom of the tube. The remaining liquid that lies above the precipitate is called a supernatant or supernate. There is a correlation between the size and density of a particle and the rate that the particle separates from a heterogeneous mixture, when the only force applied is that of gravity. The larger the size and the larger the density of the particles, the faster they separate from the mixture. By applying a larger effective gravitational force to the mixture, like a ...
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Buoyant Density Centrifugation
Buoyant density centrifugation (also isopycnic centrifugation or equilibrium density-gradient centrifugation) uses the concept of buoyancy to separate molecules in solution by their differences in density. Implementation Historically a caesium chloride (CsCl) solution was often used, but more commonly used density gradients are sucrose or Percoll. The sample is put on top of the solution, and then the tube is spun at a very high speed for an extended time, at times lasting days. The CsCl molecules become densely packed toward the bottom, so even layers of different densities form. Since the original solution was approximately the same density, they go to a level where their density and the CsCl density are the same, to which they form a sharp, distinctive band. Isotope separation This method very sharply separates molecules, and is so sharp that it can even separate different molecular isotopes from one another. DNA separation Buoyant density of majority of DNA is 1.7g/cm3 whi ...
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Gradient Centrifugation Of Infected Red Blood Cells
In vector calculus, the gradient of a scalar-valued differentiable function of several variables is the vector field (or vector-valued function) \nabla f whose value at a point p is the "direction and rate of fastest increase". If the gradient of a function is non-zero at a point , the direction of the gradient is the direction in which the function increases most quickly from , and the magnitude of the gradient is the rate of increase in that direction, the greatest absolute directional derivative. Further, a point where the gradient is the zero vector is known as a stationary point. The gradient thus plays a fundamental role in optimization theory, where it is used to maximize a function by gradient ascent. In coordinate-free terms, the gradient of a function f(\bf) may be defined by: :df=\nabla f \cdot d\bf where ''df'' is the total infinitesimal change in ''f'' for an infinitesimal displacement d\bf, and is seen to be maximal when d\bf is in the direction of the gradi ...
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Endotoxin
Lipopolysaccharides (LPS) are large molecules consisting of a lipid and a polysaccharide that are bacterial toxins. They are composed of an O-antigen, an outer core, and an inner core all joined by a covalent bond, and are found in the outer membrane of Gram-negative bacteria. Today, the term ''endotoxin'' is often used synonymously with LPS, although there are a few endotoxins (in the original sense of toxins that are inside the bacterial cell that are released when the cell disintegrates) that are not related to LPS, such as the so-called delta endotoxin proteins produced by ''Bacillus thuringiensis''. Lipopolysaccharides can have substantial impacts on human health, primarily through interactions with the immune system. LPS is a potent activator of the immune system and pyrogen (agent that causes fever). In severe cases, LPS can play a role in causing septic shock. In lower levels and over a longer time period, there is evidence LPS may play an important and harmful role i ...
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FDA Warning Letter
An FDA warning letter is an official message from the United States Food and Drug Administration (FDA) to a manufacturer or other organization that has violated some rule in a federally regulated activity. The FDA defines an FDA warning letter as: ... a correspondence that notifies regulated industry about violations that FDA has documented during its inspections or investigations. Typically, a Warning Letter notifies a responsible individual or firm that the Agency considers one or more products, practices, processes, or other activities to be in violation of the Federal Food, Drug, and Cosmetic Act (the Act), its implementing regulations and other federal statutes. Warning Letters should only be issued for violations of regulatory significance, i.e., those that may actually lead to an enforcement action if the documented violations are not promptly and adequately corrected. A Warning Letter is one of the Agency's principal means of achieving prompt voluntary compliance with t ...
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Pharmacia
Pharmacia was a pharmaceutical and biotechnological company in Sweden that merged with the American pharmaceutical company Upjohn in 1995. History Pharmacia company was founded in 1911 in Stockholm, Sweden by pharmacist Gustav Felix Grönfeldt at the ''Elgen'' Pharmacy. The company is named after the Greek word φαρμακεία, transliterated ''pharmakeia'', which means 'sorcery'. In the company's early days, much of its profits were derived from the "miracle medicine" Phospho-Energon. During World War II, Swedish chemist Björn Ingelman (who worked for Arne Tiselius at Uppsala university) researched various uses for the polysaccharide dextran. Together with the medical researcher Anders Grönwall, he discovered that dextran could be used as a replacement for blood plasma in blood transfusions, for which there could be a large need in wartime. Pharmacia, which then was still a small company, was contacted in 1943 and its CEO Elis Göth was very interested. The product Mac ...
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Intrauterine Insemination
Artificial insemination is the deliberate introduction of sperm into a female's cervix or uterine cavity for the purpose of achieving a pregnancy through in vivo fertilization by means other than sexual intercourse. It is a fertility treatment for humans, and is common practice in animal breeding, including dairy cattle (see Frozen bovine semen) and pigs. Artificial insemination may employ assisted reproductive technology, sperm donation and animal husbandry techniques. Artificial insemination techniques available include intracervical insemination (ICI) and intrauterine insemination (IUI). Humans History The first recorded case of artificial insemination was John Hunter in 1790, who helped impregnate a linen draper's wife. The first reported case of artificial insemination by donor occurred in 1884: William H. Pancoast, a professor in Philadelphia, took sperm from his "best looking" student to inseminate an anesthetized woman without her knowledge. The case was repor ...
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In Vitro Fertilization
In vitro fertilisation (IVF) is a process of fertilisation where an egg is combined with sperm in vitro ("in glass"). The process involves monitoring and stimulating an individual's ovulatory process, removing an ovum or ova (egg or eggs) from their ovaries and letting sperm fertilise them in a culture medium in a laboratory. After the fertilised egg (zygote) undergoes embryo culture for 2–6 days, it is transferred by catheter into the uterus, with the intention of establishing a successful pregnancy. IVF is a type of assisted reproductive technology used for infertility treatment, gestational surrogacy, and, in combination with pre-implantation genetic testing, avoiding transmission of genetic conditions. A fertilised egg from a donor may implant into a surrogate's uterus, and the resulting child is genetically unrelated to the surrogate. Some countries have banned or otherwise regulate the availability of IVF treatment, giving rise to fertility tourism. Restrictions on ...
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Semen
Semen, also known as seminal fluid, is an organic bodily fluid created to contain spermatozoa. It is secreted by the gonads (sexual glands) and other sexual organs of male or hermaphroditic animals and can fertilize the female ovum. Semen is produced and originates from the seminal vesicle, which is located in the pelvis. The process that results in the discharge of semen from the urethral orifice is called ejaculation. In humans, seminal fluid contains several components besides spermatozoa: proteolytic and other enzymes as well as fructose are elements of seminal fluid which promote the survival of spermatozoa, and provide a medium through which they can move or "swim". The fluid is designed to be discharged deep into the vagina, so the spermatozoa can pass into the uterus and form a zygote with an egg. Semen is also a form of genetic material. In animals, semen has been collected for cryoconservation. Cryoconservation of animal genetic resources is a practice ...
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Sperm
Sperm is the male reproductive cell, or gamete, in anisogamous forms of sexual reproduction (forms in which there is a larger, female reproductive cell and a smaller, male one). Animals produce motile sperm with a tail known as a flagellum, which are known as spermatozoa, while some red algae and fungi produce non-motile sperm cells, known as spermatia. Flowering plants contain non-motile sperm inside pollen, while some more basal plants like ferns and some gymnosperms have motile sperm. Sperm cells form during the process known as spermatogenesis, which in amniotes (reptiles and mammals) takes place in the seminiferous tubules of the testes. This process involves the production of several successive sperm cell precursors, starting with spermatogonia, which differentiate into spermatocytes. The spermatocytes then undergo meiosis, reducing their chromosome number by half, which produces spermatids. The spermatids then mature and, in animals, construct a tail, or fla ...
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Assisted Reproductive Technology
Assisted reproductive technology (ART) includes medical procedures used primarily to address infertility. This subject involves procedures such as in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), cryopreservation of gametes or embryos, and/or the use of fertility medication. When used to address infertility, ART may also be referred to as fertility treatment. ART mainly belongs to the field of reproductive endocrinology and infertility. Some forms of ART may be used with regard to fertile couples for genetic purpose (see preimplantation genetic diagnosis). ART may also be used in surrogacy arrangements, although not all surrogacy arrangements involve ART. The existence of sterility will not always require ART to be the first option to consider, as there are occasions when its cause is a mild disorder that can be solved with more conventional treatments or with behaviors based on promoting health and reproductive habits. Procedures General With ART, ...
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Osmotic Concentration
Osmotic concentration, formerly known as osmolarity, is the measure of solute concentration, defined as the number of osmoles (Osm) of solute per litre (L) of solution (osmol/L or Osm/L). The osmolarity of a solution is usually expressed as Osm/L (pronounced "osmolar"), in the same way that the molarity of a solution is expressed as "M" (pronounced "molar"). Whereas molarity measures the number of moles of solute per unit volume of solution, osmolarity measures the number of ''osmoles of solute particles'' per unit volume of solution. This value allows the measurement of the osmotic pressure of a solution and the determination of how the solvent will diffuse across a semipermeable membrane (osmosis) separating two solutions of different osmotic concentration. Unit The unit of osmotic concentration is the osmole. This is a non- SI unit of measurement that defines the number of moles of solute that contribute to the osmotic pressure of a solution. A milliosmole (mOsm) is 1/1,00 ...
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