The ultracentrifuge is a centrifuge optimized for spinning a rotor at very high speeds, capable of generating acceleration as high as 1,000,000 g (9,800 km/s²). There are two kinds of ultracentrifuges, the preparative and the analytical ultracentrifuge. Both classes of instruments find important uses in molecular biology, biochemistry and polymer science. Theodor Svedberg invented the analytical ultracentrifuge in 1923, and won the Nobel Prize in Chemistry in 1926 for his research on colloids and proteins using the ultracentrifuge.
The vacuum ultracentrifuge was invented by Edward Greydon Pickels. It was his contribution of the vacuum which allowed a reduction in friction generated at high speeds. Vacuum systems also enabled the maintenance of constant temperature.
In 1946, Pickels cofounded Spinco (Specialized Instruments Corp.) and marketed an ultracentrifuge based on his design. Pickels, however, considered his design to be complicated and developed a more “foolproof” version. But even with the enhanced design, sales of the technology remained low, and Spinco almost went bankrupt. The company survived and was the first to commercially manufacture ultracentrifuges, in 1947. In 1949, Spinco introduced the Model L, the first preparative ultracentrifuge to reach a maximum speed of 40,000 rpm. In 1954, Beckman Instruments (now Beckman Coulter) purchased the company, forming the basis of its Spinco centrifuge division.
Sedimentation velocity experiments aim to interpret the entire time-course of sedimentation, and report on the shape and molar mass of the dissolved macromolecules, as well as their size-distribution. The size resolution of this method scales approximately with the square of the particle radii, and by adjusting the rotor speed of the experiment size-ranges from 100 Da to 10 GDa can be covered. Sedimentation velocity experiments can also be used to study reversible chemical equilibria between macromolecular species, by either monitoring the number and molar mass of macromolecular complexes, by gaining information about the complex composition from multi-signal analysis exploiting differences in each components spectroscopic signal, or by following the composition dependence of the sedimentation rates of the macromolecular system, as described in Gilbert-Jenkins theory.
Sedimentation equilibrium experiments are concerned only with the final steady-state of the experiment, where sedimentation is balanced by diffusion opposing the concentration gradients, resulting in a time-independent concentration profile. Sedimentation equilibrium distributions in the centrifugal field are characterized by Boltzmann distributions. This experiment is insensitive to the shape of the macromolecule, and directly reports on the molar mass of the macromolecules and, for chemically reacting mixtures, on chemical equilibrium constants.
The kinds of information that can be obtained from an analytical ultracentrifuge include the gross shape of macromolecules, the conformational changes in macromolecules, and size distributions of macromolecular samples. For macromolecules, such as proteins, that exist in chemical equilibrium with different non-covalent complexes, the number and subunit stoichiometry of the complexes and equilibrium constant constants can be studied.
Preparative rotors are used in biology for pelleting of fine particulate fractions, such as cellular organelles (mitochondria, microsomes, ribosomes) and viruses. They can also be used for gradient 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.
Contract Notice: Department of Veterans Affairs (Indiana) Issues Solicitation for "Beckman Coulter Optima XE 90K Ultracentrifuge & Accessories"
Apr 07, 2012; WASHINGTON, April 11 -- Department of Veterans Affairs, Indianapolis VAMC has a requirement for "Beckman Coulter Optima XE 90K...