An ultracentrifuge is a
centrifuge
A centrifuge is a device that uses centrifugal force to separate various components of a fluid. This is achieved by spinning the fluid at high speed within a container, thereby separating fluids of different densities (e.g. cream from milk) or ...
optimized for spinning a rotor at very high speeds, capable of generating acceleration as high as (approx. ). There are two kinds of ultracentrifuges, the preparative and the analytical ultracentrifuge. Both classes of instruments find important uses in
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 physi ...
,
biochemistry
Biochemistry or biological chemistry is the study of chemical processes within and relating to living organisms. A sub-discipline of both chemistry and biology, biochemistry may be divided into three fields: structural biology, enzymology and ...
, and
polymer
A polymer (; Greek '' poly-'', "many" + ''-mer'', "part")
is a substance or material consisting of very large molecules called macromolecules, composed of many repeating subunits. Due to their broad spectrum of properties, both synthetic a ...
science.
[Susan R. Mikkelsen & Eduardo Cortón. Bioanalytical Chemistry, Ch. 13. Centrifugation Methods. John Wiley & Sons, Mar 4, 2004, pp. 247-267.]
History
In 1924
Theodor Svedberg
Theodor Svedberg (30 August 1884 – 25 February 1971) was a Swedish chemist and Nobel laureate for his research on colloids and proteins using the ultracentrifuge. Svedberg was active at Uppsala University from the mid 1900s to late 1940s. ...
built a centrifuge capable of generating 7,000 g (at 12,000 rpm), and called it the ultracentrifuge, to juxtapose it with the
Ultramicroscope
An ultramicroscope is a microscope with a system that lights the object in a way that allows viewing of tiny particles via light scattering, and not light reflection or absorption. When the diameter of a particle is below or near the wavelength o ...
that had been developed previously. In 1925-1926 Svedberg constructed a new ultracentrifuge that permitted fields up to 100,000 g (42,000 rpm).
Modern ultracentrifuges are typically classified as allowing greater than 100,000 g. Svedberg won the
Nobel Prize in Chemistry
)
, image = Nobel Prize.png
, alt = A golden medallion with an embossed image of a bearded man facing left in profile. To the left of the man is the text "ALFR•" then "NOBEL", and on the right, the text (smaller) "NAT•" then "M ...
in 1926 for his research on colloids and proteins using the ultracentrifuge.
[Joe Rosen; Lisa Quinn Gothard. ]
Encyclopedia of Physical Science
'. Infobase Publishing; 2009. . p. 77.
The vacuum ultracentrifuge was invented by
Edward Greydon Pickels in the Physics Department at the
University of Virginia
The University of Virginia (UVA) is a public research university in Charlottesville, Virginia. Founded in 1819 by Thomas Jefferson, the university is ranked among the top academic institutions in the United States, with highly selective ad ...
. It was his contribution of the
vacuum
A vacuum is a space devoid of matter. The word is derived from the Latin adjective ''vacuus'' for "vacant" or " void". An approximation to such vacuum is a region with a gaseous pressure much less than atmospheric pressure. Physicists often di ...
which allowed a reduction in
friction
Friction is the force resisting the relative motion of solid surfaces, fluid layers, and material elements sliding against each other. There are several types of friction:
*Dry friction is a force that opposes the relative lateral motion of ...
generated at high speeds. Vacuum systems also enabled the maintenance of constant
temperature
Temperature is a physical quantity that expresses quantitatively the perceptions of hotness and coldness. Temperature is measurement, measured with a thermometer.
Thermometers are calibrated in various Conversion of units of temperature, temp ...
across the sample, eliminating
convection current
Convection is single or multiphase fluid flow that occurs spontaneously due to the combined effects of material property heterogeneity and body forces on a fluid, most commonly density and gravity (see buoyancy). When the cause of the conve ...
s that interfered with the interpretation of sedimentation results.
[Elzen B. ''Vacuum ultracentrifuge.'' In: Encyclopedia of 20th-Century Technology, Colin Hempstead & William Worthington, eds. Routledge, 2005. p. 868.]
In 1946, Pickels cofounded
Spinco (Specialized Instruments Corp.) to market analytical and preparative ultracentrifuges based on his design. Pickels considered his design to be too complicated for commercial use and developed a more easily operated, “foolproof” version. But even with the enhanced design, sales of analytical centrifuges remained low, and Spinco almost went bankrupt. The company survived by concentrating on sales of preparative ultracentrifuge models, which were becoming popular as workhorses in biomedical laboratories.
[ In 1949, Spinco introduced the Model L, the first preparative ultracentrifuge to reach a maximum speed of 40,000 ]rpm
Revolutions per minute (abbreviated rpm, RPM, rev/min, r/min, or with the notation min−1) is a unit of rotational speed or rotational frequency for rotating machines.
Standards
ISO 80000-3:2019 defines a unit of rotation as the dimensionl ...
. In 1954, Beckman Instruments
Beckman Coulter Inc. is a Danaher Corporation company that develops, manufactures, and markets products that simplify, automate and innovate complex biomedical testing. It operates in two industries: Diagnostics and Life Sciences. For more than ...
(later Beckman Coulter
Beckman Coulter Inc. is a Danaher Corporation company that develops, manufactures, and markets products that simplify, automate and innovate complex biomedical testing. It operates in two industries: Diagnostics and Life Sciences. For more than ...
) purchased the company, forming the basis of its Spinco centrifuge division.
Instrumentation
Ultracentrifuges are available with a wide variety of rotors suitable for a great range of experiments. Most rotors are designed to hold tubes that contain the samples. ''Swinging bucket rotors'' allow the tubes to hang on hinges so the tubes reorient to the horizontal as the rotor initially accelerate. ''Fixed angle rotors'' are made of a single block of material and hold the tubes in cavities bored at a predetermined angle. ''Zonal rotors'' are designed to contain a large volume of sample in a single central cavity rather than in tubes. Some zonal rotors are capable of dynamic loading and unloading of samples while the rotor is spinning at high speed.
Preparative rotors are used in biology for pelleting of fine particulate fractions, such as cellular organelles ( mitochondria, microsome
In cell biology, microsomes are heterogeneous vesicle-like artifacts (~20-200 nm diameter) re-formed from pieces of the endoplasmic reticulum (ER) when eukaryotic cells are broken-up in the laboratory; microsomes are not present in healthy, liv ...
s, ribosomes) and virus
A virus is a submicroscopic infectious agent that replicates only inside the living cells of an organism. Viruses infect all life forms, from animals and plants to microorganisms, including bacteria and archaea.
Since Dmitri Ivanovsk ...
es. They can also be used for gradient
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 gr ...
separations, in which the tubes are filled from top to bottom with an increasing concentration of a dense substance in solution. Sucrose gradients are typically used for separation of cellular organelles. Gradients of caesium salts are used for separation of nucleic acids. After the sample has spun at high speed for sufficient time to produce the separation, the rotor is allowed to come to a smooth stop and the gradient is gently pumped out of each tube to isolate the separated components.
Hazards
The tremendous rotational kinetic energy
In physics, the kinetic energy of an object is the energy that it possesses due to its motion.
It is defined as the work needed to accelerate a body of a given mass from rest to its stated velocity. Having gained this energy during its acc ...
of the rotor in an operating ultracentrifuge makes the catastrophic failure of a spinning rotor a serious concern, as it can explode spectacularly. Rotors conventionally have been made from high strength-to-weight metals such as aluminum or titanium. The stresses of routine use and harsh chemical solutions eventually cause rotors to deteriorate. Proper use of the instrument and rotors within recommended limits and careful maintenance of rotors to prevent corrosion and to detect deterioration is necessary to mitigate this risk.
More recently some rotors have been made of lightweight carbon fiber composite material, which are up to 60% lighter, resulting in faster acceleration/deceleration rates. Carbon fiber composite rotors also are corrosion-resistant, eliminating a major cause of rotor failure.[Piramoon, Sheila. "Carbon fibers boost centrifuge flexibility: advancements in centrifuge rotors over the years have led to improved lab productivity." Laboratory Equipment Mar. 2011: 12+. General Reference Center GOLD. Web. 15 Feb. 2015.]
See also
*Analytical ultracentrifugation Analytical ultracentrifugation is an analytical technique which combines an ultracentrifuge with optical monitoring systems.
In an analytical ultracentrifuge (commonly abbreviated as AUC), a sample’s sedimentation profile is monitored in real tim ...
*Gas centrifuge
A gas centrifuge is a device that performs isotope separation of gases. A centrifuge relies on the principles of centrifugal force accelerating molecules so that particles of different masses are physically separated in a gradient along the radiu ...
*Theodor Svedberg
Theodor Svedberg (30 August 1884 – 25 February 1971) was a Swedish chemist and Nobel laureate for his research on colloids and proteins using the ultracentrifuge. Svedberg was active at Uppsala University from the mid 1900s to late 1940s. ...
*Differential centrifugation
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 o ...
*Buoyant density ultracentrifugation
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 c ...
*Zippe-type centrifuge
The Zippe-type centrifuge is a gas centrifuge designed to enrich the rare fissile isotope uranium-235 (235U) from the mixture of isotopes found in naturally occurring uranium compounds. The isotopic separation is based on the slight difference in ...
References
External links
07702/pdf Modern Analytical Ultracentrifugation in Protein Science: A tutorial review
Analytical Ultracentrifugation as a Contemporary Biomolecular Research Tool.
Multi-signal analysis
Gilbert-Jenkins theory
{{Authority control
Centrifuges