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Sucrose density gradient centrifugation for efficient separation of engineered nanoparticles from a model organism, Caenorhabditis elegans

Published

Author(s)

Monique E. Johnson, Antonio R. Montoro Bustos, Elijah J. Petersen, Shannon K. Hanna, Karen E. Murphy, Lee L. Yu, Bryant C. Nelson, Michael R. Winchester

Abstract

This protocol describes a strategy for separating Caenorhabditis elegans (C. elegans) from engineered nanomaterials, a critical step for accurate quantification in uptake studies. Conventional rinsing procedures, typically comprised of water or buffer rinsing steps followed by centrifugation, have been previously tested and deemed unsuitable for this purpose, even at environmentally relevant exposure concentrations. Therefore, this protocol details the development and assessment of a unique sucrose gradient density centrifugation procedure for the efficient separation of nanoparticles and C. elegans at the end of an exposure protocol. The example outlined in this SP pertains to the measurement of gold nanoparticles (AuNPs) and C. elegans. However, this method can potentially be implemented in bioaccumulation studies involving other organisms and with different metallonanoparticles. Visit http://nist.gov/mml/np-measurement-protocols.cfm to check for revisions and additions to this protocol or for new protocols in the series. We also encourage users to report citations to published work in which this protocol has been applied.
Citation
Special Publication (NIST SP) - 1200-24
Report Number
1200-24

Keywords

sucrose density gradient separation, Caenorhabditis elegans, gold nanoparticles
Created October 11, 2017, Updated November 10, 2018