Enhanced Sample Handling for Analytical Ultracentrifugation with 3D-Printed Centerpieces

Samuel C. To, Chad A. Brautigam, Sumit K. Chaturvedi, Mary T. Bollard, Jonathan Krynitsky, John W. Kakareka, Thomas J. Pohida, Huaying Zhao, Peter Schuck

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

The centerpiece of the sample cell assembly in analytical ultracentrifugation holds the sample solution between windows, sealed against high vacuum, and is shaped such that macromolecular migration in centrifugal fields exceeding 200 000g can proceed undisturbed by walls or convection while concentration profiles are imaged with optical detection systems aligned perpendicular to the plane of rotation. We have recently shown that 3D printing using various materials allows inexpensive and rapid manufacturing of centerpieces. In the present work, we expand this endeavor to examine the accuracy of the measured sedimentation process, as well as short-term durability of the centerpieces. We find that 3D-printed centerpieces can be used many times and can provide data equivalent in quality to commonly used commercial epoxy resin centerpieces. Furthermore, 3D printing enables novel designs adapted to particular experimental objectives because they offer unique opportunities, for example, to create well-defined curved surfaces, narrow channels, and embossed features. We present examples of centerpiece designs exploiting these capabilities for improved AUC experiments. This includes narrow sector centerpieces that substantially reduce the required sample volume while maintaining the standard optical path length; thin centerpieces with integrated window holders to provide very short optical pathlengths that reduce optical aberrations at high macromolecular concentrations; long-column centerpieces that increase the observable distance of macromolecular migration for higher-precision sedimentation coefficients; and three-sector centerpieces that allow doubling the number of samples in a single run while reducing the sample volumes. We find each of these designs allows unimpeded macromolecular sedimentation and can provide high-quality sedimentation data.

Original languageEnglish (US)
Pages (from-to)5866-5873
Number of pages8
JournalAnalytical chemistry
Volume91
Issue number9
DOIs
StatePublished - May 7 2019

ASJC Scopus subject areas

  • Analytical Chemistry

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    To, S. C., Brautigam, C. A., Chaturvedi, S. K., Bollard, M. T., Krynitsky, J., Kakareka, J. W., Pohida, T. J., Zhao, H., & Schuck, P. (2019). Enhanced Sample Handling for Analytical Ultracentrifugation with 3D-Printed Centerpieces. Analytical chemistry, 91(9), 5866-5873. https://doi.org/10.1021/acs.analchem.9b00202