AN IMPROVED AIR-DRIVEN TYPE OF ULTRACENTRIFUGE FOR MOLECULAR SEDIMENTATION

Abstract

1. A description is given of the construction details and operation characteristics of an improved type of air-driven ultracentrifuge operating in vacuum and suitable for the determination of sedimentation constants of protein molecules. 2. The rotor of the centrifuge is made of a forged aluminum alloy; it is oval in shape, measures 185 mm. at its greatest diameter, and weighs 3,430 gm. It carries a transparent cell located at a distance of 65 mm. from the axis of rotation and designed to accommodate a fluid column 15 mm. high. 3. The rotor has been run repeatedly over long periods at a speed of 60,000 R.P.M., which corresponds to a centrifugal force of 260,000 times gravity in the center of the cell. At this speed no deformation of the rotor nor leakage of the cell has been observed. 4. The sharp definition of sedimentation photographs taken at high speed serves to indicate the absence of detectable vibrations in the centrifuge. 5. When a vacuum of less than 1 micron of mercury is maintained in the centrifuge chamber, the rise in the rotor temperature amounts to only 1 or 2 degrees C. after several hours' run at high speed. 6. There has been no evidence of convection currents interfering with normal sedimentation of protein molecules in the centrifugal field. 7. A driving air pressure of about 18 pounds per square inch is sufficient to maintain the centrifuge at a steady speed of 60,000 R.P.M. With a driving pressure of 80 pounds per square inch, it can be accelerated to this speed in less than 20 minutes, and also brought to rest in about the same length of time by the application of the braking system. 8. The adaptation of Svedberg's optical systems to this centrifuge for photographically recording the movement of sedimentation boundaries is described.