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Centrifugation

Centrifugation is a process that uses centrifugal force to separate mixtures by spinning samples in a centrifuge. It increases the effective gravitational force on a mixture in a test tube, rapidly bringing denser precipitates to the bottom. The remaining supernatant liquid is then separated from the precipitate. Differential centrifugation is a technique used to separate particles like cells and organelles based on size or density differences by spinning samples through solutions with varying densities. While effective, density gradient centrifugation in tubes is limited by the small sample sizes that can be processed, requiring many tubes for large quantities. Zonal rotors allow for fractionating much larger sample volumes.

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43 views2 pages

Centrifugation

Centrifugation is a process that uses centrifugal force to separate mixtures by spinning samples in a centrifuge. It increases the effective gravitational force on a mixture in a test tube, rapidly bringing denser precipitates to the bottom. The remaining supernatant liquid is then separated from the precipitate. Differential centrifugation is a technique used to separate particles like cells and organelles based on size or density differences by spinning samples through solutions with varying densities. While effective, density gradient centrifugation in tubes is limited by the small sample sizes that can be processed, requiring many tubes for large quantities. Zonal rotors allow for fractionating much larger sample volumes.

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rockingtwo07
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Centrifugation

Centrifugation is a process that involves the use of the centrifugal force for the
separation of mixtures, used in industry and in laboratory settings. In chemistry and biology,
centrifugation increases the effective gravitational force on a mixture in a test tube, to rapidly
and completely bring the precipitate ("pellet") to the bottom of the tube. The
remaining solution is called the "supernate," "supernatant," or supernatant liquid. The
supernatant liquid is then separated from the precipitate by decantation or withdrawal with a
Pasteur pipette.
The equipment used for centrifugation is called a centrifuge, and the vessel that spins the
samples is called a rotor. Generally, a motor causes the rotor to spin around a fixed axis,
applying a force perpendicular to the axis. The centrifuge works using the sedimentation
principle, where the centripetal acceleration is used to separate substances of greater and lesser
density.
There are many different kinds of centrifuges, including those for very specialized
purposes. In the chemical and food industries, special centrifuges can process a continuous
stream of particle-laden liquid.
English military engineer Laval (1707-1751) invented a whirling arm apparatus to
determine drag, and Antonin Prandl invented the first centrifuge in order to separate cream from
milk to make it easier to churn butter.
Differential Centrifugation
If you had sufficient time and a vibration-free environment, you could patiently wait and
the force of gravity would bring most suspended particles to the bottom of a centrifuge tube. The
smallest particles would probably stay in suspension due to brownian motion, and most
macromolecules would be uniformly distributed because they would be in solution rather than
suspension. I don't know about you, but I don't have the kind of patience needed in order to rely
solely on gravity for separation of solid from liquid components. Besides, for practical purposes
the pellet you obtained would be way too easily disrupted for effective separation of solid
material from supernatant. Gravity would not be a terribly effective way of separating suspended
materials based on size or other characteristics.
Density gradient centrifugation using tubes is the most widely employed technique for
separating cells and cell organelles and for isolating cellular macromolecules. However, although
it is one of the cell biologists most valuable tools, it is not without disadvantages, as the amount
of material that can be fractionated in a single tube is so small.

When large quantities of sample must be fractionated (to isolate sparse organelles such as
lysosomes or peroxisomes), a very large number of tubes and gradients is needed. Much larger
quantities of sample may be fractionated using zonal rotors.

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