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Homogenization, in
Homogenization, in cell biology
Cell biology (also cellular biology or cytology) is a branch of biology that studies the structure, function, and behavior of cells. All living organisms are made of cells. A cell is the basic unit of life that is responsible for the living an ...
or molecular biology
Molecular biology is the branch of biology that seeks to understand the molecular basis of biological activity in and between cells, including biomolecular synthesis, modification, mechanisms, and interactions. The study of chemical and phys ...
, is a process whereby different fractions of a biological sample become equal in composition. It can be a disease sign in histopathology
Histopathology (compound of three Greek words: ''histos'' "tissue", πάθος ''pathos'' "suffering", and -λογία '' -logia'' "study of") refers to the microscopic examination of tissue in order to study the manifestations of disease. Sp ...
, 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
The enzyme-linked immunosorbent assay (ELISA) (, ) is a commonly used analytical biochemistry assay, first described by Eva Engvall and Peter Perlmann in 1971. The assay uses a solid-phase type of enzyme immunoassay (EIA) to detect the presen ...
and western blot.
Methods
Homogenization of tissue in solution is often performed simultaneously with celllysis
Lysis ( ) is the breaking down of the membrane of a cell, often by viral, enzymic, or osmotic (that is, "lytic" ) mechanisms that compromise its integrity. A fluid containing the contents of lysed cells is called a ''lysate''. In molecular b ...
. To prevent lysis however, the tissue (or collection of cells, e.g. from cell culture
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 tec ...
) can be kept at temperatures slightly above zero to prevent autolysis, and in an isotonic solution to prevent osmotic
Osmosis (, ) is the spontaneous net movement or diffusion of solvent molecules through a selectively-permeable membrane from a region of high water potential (region of lower solute concentration) to a region of low water potential (region ...
damage.
If freezing the tissue is possible, cryohomogenization can be performed under "dry" conditions, and is often the method of choice whenever it is desirable to collect several distinct molecular classes (e.g. both protein
Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residues. Proteins perform a vast array of functions within organisms, including catalysing metabolic reactions, DNA replication, res ...
and RNA) from a single sample, or combined set of samples, or when long-term storage of part of the sample is desired. Cryohomogenization can be carried out using a supercooled mortar and pestle (classic approach), or the tissue can be homogenized by crushing it into a fine powder inside a clean plastic bag resting against a supercooled solid metal block (more recently developed and more efficient technique).
High-pressure homogenization is used to isolate the contents of Gram-positive bacteria
In bacteriology, gram-positive bacteria are bacteria that give a positive result in the Gram stain test, which is traditionally used to quickly classify bacteria into two broad categories according to their type of cell wall.
Gram-positive bac ...
, since these cells are exceptionally resistant to lysis, and may be combined with high-temperature sterilization.
Dounce homogenization is a technique suitable for soft mammalian tissues, while lysis of mammalian cells has also been demonstrated via centrifugation.
References
{{DEFAULTSORT:Homogenization (Biology) Cell anatomy Cell biology Laboratory techniques