Collecting and processing records from the ultracentrifuge in “Real-Time” using an on-line computer

• Published in: Biophysical chemistry 1976-07, Vol.5 (1), p.43-53
• Main Authors: Spragg, S.P., Barnett, W.A., Wilcox, J.K., Roche, J.
• Format: Article
• Language: English
• Subjects: Computers, Hybrid; Macromolecular Substances; Mathematics; Molecular Weight; Online Systems; Serum Albumin, Bovine; Ultracentrifugation - instrumentation; Ultracentrifugation - methods
• ISSN: 0301-4622; 1873-4200
• DOI: 10.1016/0301-4622(76)80025-7

An analytical system consisting of an analytical centrifuge coupled ‘on-line’ to a computer was assembled and tested. Collection of records from up to 9 solutions was achieved through programmes which sum readings to reduce noise as well as controlling the positioning of the scanner. With this system it was found that the limit on accuracy for molecular weights at concentrations less than 0.01 g cm −3 was ± 3% estimated from sedimentation equilibrium experiments. The Same system was used to collect records for similar concentrations from velocity experiments by employing a scanning schlicren. In this case the accuracy in estimating sedimentation coefficients was similar to those found when measuring photographs. Since the collection yields detailed information about the shape of the sedimenting boundary, the centroids of the boundary were routinely computed by second moment analysis rather than relying on the position of the maximum of the schlieren peak. In the same analysis estimates of diffusion coefficients were made routinely by calculating corrected height/area ratios for each scan. These calculations were made during the real-time of the experiment, so making available molecular parameters rather than records which must be evaluated some time after stopping the experiment.