Back طرد مركزي تبايني Arabic Centrifugació diferencial Catalan Differentielle Zentrifugation German گریز از مرکز تفاضلی Persian Centrifugation analytique French Sentrifugasi diferensial ID Диференційне центрифугування Ukrainian
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In biochemistry and cell biology, differential centrifugation (also known as differential velocity centrifugation) is a common procedure used to separate organelles and other sub-cellular particles based on their sedimentation rate. Although often applied in biological analysis, differential centrifugation is a general technique also suitable for crude purification of non-living suspended particles (e.g. nanoparticles, colloidal particles, viruses). In a typical case where differential centrifugation is used to analyze cell-biological phenomena (e.g. organelle distribution), a tissue sample is first lysed to break the cell membranes and release the organelles and cytosol. The lysate is then subjected to repeated centrifugations, where particles that sediment sufficiently quickly at a given centrifugal force for a given time form a compact "pellet" at the bottom of the centrifugation tube.[1]
After each centrifugation, the supernatant (non-pelleted solution) is removed from the tube and re-centrifuged at an increased centrifugal force and/or time. Differential centrifugation is suitable for crude separations on the basis of sedimentation rate, but more fine grained purifications may be done on the basis of density through equilibrium density-gradient centrifugation.[2] Thus, the differential centrifugation method is the successive pelleting of particles from the previous supernatant, using increasingly higher centrifugation forces.[3] Cellular organelles separated by differential centrifugation maintain a relatively high degree of normal functioning, as long as they are not subject to denaturing conditions during isolation.[4][5]
- ^ Ohlendieck, Kay; Harding, Stephen E. (19 April 2018). "Centrifugation and Ultracentrifugation". Wilson and Walker's Principles and Techniques of Biochemistry and Molecular Biology: 424–453. doi:10.1017/9781316677056.014. ISBN 9781107162273.
- ^ Darnell, James; Baltimore, David; Matsudaira, Paul; Zipursky, S. Lawrence; Berk, Arnold; Lodish, Harvey (2000). "Purification of Cells and Their Parts".
{{cite journal}}: Cite journal requires|journal=(help) - ^ Griffith, Owen Mitch (2010). Practical Techniques for Centrifugal Separations – Application Guide (PDF). Principles & Techniques of Biochemistry and Molecular Biology. pp. 1–27. Archived from the original (PDF) on 2023-02-07. Retrieved 2020-10-14.
- ^ Gerald Karp (19 October 2009). Cell and Molecular Biology: Concepts and Experiments. John Wiley & Sons. pp. 28–. ISBN 978-0-470-48337-4.
- ^ Livshits, Mikhail A.; Khomyakova, Elena; Evtushenko, Evgeniy G.; Lazarev, Vassili N.; Kulemin, Nikolay A.; Semina, Svetlana E.; Generozov, Edward V.; Govorun, Vadim M. (30 November 2015). "Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol". Scientific Reports. 5 (1): 17319. Bibcode:2015NatSR...517319L. doi:10.1038/srep17319. ISSN 2045-2322. PMC 4663484. PMID 26616523. S2CID 14200669.