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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.
History
By 1923 Theodor Svedberg and his student H. Rinde had successfully analyzed large-grained sols
in terms of their gravitational sedimentation. Sols consist of a substance evenly distributed in another
substance, also known as a colloid. However, smaller grained sols, such as those containing gold, could
not be analyzed. To investigate this problem Svedberg developed an analytical centrifuge, equipped with
a photographic absorption system, which would exert a much greater centrifugal effect. In addition, he
developed the theory necessary to measure molecular weight. During this time, Svedberg’s attention
shifted from gold to proteins.
By 1900, it was generally accepted that proteins were composed of amino acids; however,
whether proteins were colloids or macromolecules was still under debate.
One protein being investigated at the time was hemoglobin. It was determined to have 712
carbon, 1,130 hydrogen, 243 oxygen, two sulfur atoms, and at least one iron atom. This gave hemoglobin
a resulting weight of approximately 16,000 Da but it was uncertain whether this value was a multiple of
one or four (dependent upon the number of iron atoms present).

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

The entire chamber is used during centrifugation and is loaded with a single density gradient. Density gradient centrifugation using tubes is the most widely employed technique for separating cells and cell organelles and for isolating cellular macromolecules. 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.and fatigue-resistance provides unequaled durability  Lightweight for simple handling and convenience  Ultimate sample temperature control for reliable results  Increased productivity with higher G-forces Types of Rotor Centrifuges Swing-Bucket Rotors  A swing-bucket rotor usually supports samples ranging in volume from 36 mL to 2. each sector-shaped compartment serving as a large centrifuge tube. it is not without disadvantages. Swingbuckets can support two types of separations: rate-zonal and isopycnic. Be sure to check if your intended rotor is compatible with your centrifuge’s manufacturer. and most macromolecules would be uniformly distributed because they would be in solution rather than suspension. However. Fixed-angle centrifuges hold the sample containers at a constant angle relative to the central axis and are used primarily for differential centrifugation. for practical purposes the pellet you obtained would be way too easily disrupted for effective separation of solid material from supernatant. Much larger quantities of sample may be fractionated using zonal rotors. a very large number of tubes and gradients is needed. The large chamber capacity of these rotors (typically 1 and 2 liters) elimi nates the need for multiple runs and multiple density gradients. . A zonal rotor consists of a large cylindrical chamber subdivided into a number of sector-shaped compartments by vertical septa (or vanes) that radiate from the axial core to the rotor wall. exchangeable and allow access for cleaning. swinging bucket and continuous flow. because the distance between the outside of the meniscus and the outside of the bottom of the tube is long enough for separation to occur. Swing-buckets are preferred for rate-zonal separations. Choice of centrifuge rotor and lid will depend on application. have a hinge where the sample containers are attached to the central rotor and are used primarily for gradient work. Fiberlite Rotors: Advanced Carbon Fiber Centrifuge Rotors  Corrosion. Swinging head (or swinging bucket) centrifuges. I don't know about you. Continuous flow centrifuges don't have individual sample vessels and are used for large volume batch separations. When large quantities of sample must be fractionated (to isolate sparse organelles such as lysosomes or peroxisomes).would probably stay in suspension due to brownian motion.2 mL. although it is one of the cell biologist’s most valuable tools. Centrifuges can be divided into types based on their rotor design: fixed angle. Look for rotors that are easily attached. Gravity would not be a terribly effective way of separating suspended materials based on size or other characteristics. as the amount of material that can be fractionated in a single tube is so small.

capacity. which makes this type of separation impractical. Separation techniques There are three types of isotope separation techniques:  Those based directly on the atomic weight of the isotope. biochemistry. the preparative and the analytical ultracentrifuge. or to collect the pellet. isotopes of the same element have nearly identical chemical properties. running speeds. .Fixed-Angle Rotors  Fixed-angle rotors are usually used for pelleting applications to either pellet particles from a suspension and remove the excess debris. biology. The most important aspect in deciding to use a fixed-angle rotor is the K factor. This process is a crucial one in the manufacture of uranium fuel for nuclear power stations. Laboratory centrifuges often can accept an range of different fixed-angle and swinging bucket rotors able to carry different numbers of centrifuge tubes and rated for specific maximum speeds.2 mL to 1 mL. capable of generating acceleration as high as 2 000 000 g (approx. Zonal rotors and continuous flow systems are capable of handing bulk and larger sample volumes. They vary widely in speed. Ultracentrifuges spin the rotors under vacuum. The use of the nuclides produced is various. By tonnage. biochemistry and clinical medicine for isolating and separating suspensions and immiscible liquids. The lower the K factor. 19 600 km/s²). Rotor cavities range from 0. The largest variety is used in research (e. respectively. the higher the pelleting efficiency. except for separation of deuterium. Ultracentrifuge: The ultracentrifuge is a centrifuge optimized for spinning a rotor at very high speeds. eliminating air resistance and enabling exact temperature control. in a laboratory-scale instrument. and other characteristics.  Those based on the small differences in chemical reaction rates produced by different atomic weights. Both classes of instruments find important uses in molecular biology.g. and polymer science. which must be operated in such a way as to produce plutonium already of suitable isotopic mix or grade. Plutonium-based weapons use plutonium produced in a nuclear reactor. separating natural uranium into enriched uranium and depleted uranium is the largest application. The K factor indicates how efficient the rotor can pellet at maximum speed. A wide variety of laboratory-scale centrifuges are used in chemistry. Controls vary from simple electrical timers to programmable models able to control acceleration and deceleration rates.There are two kinds of ultracentrifuges. Isotope separation Isotope separation is the process of concentrating specific isotopes of a chemical element by removing other isotopes. temperature control. in chemistry where atoms of "marker" nuclide are used to figure out reaction mechanisms). and temperature regimes. and is also required for the creation of uranium based nuclear weapons. While different chemical elements can be purified through chemical processes.

. such as nuclear resonances. Those based on properties not directly connected to atomic weight.