UNIT 8.5 Centrifugal Elutriation to Obtain Synchronous Populations of Cells
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Cell Biology
How to Cite
Wahl, A. F. and Donaldson, K. L. 2001. Centrifugal Elutriation to Obtain Synchronous Populations of Cells. Current Protocols in Cell Biology. 2:8.5:8.5.1–8.5.16.
- Published Online: 1 MAY 2001
- Published Print: APR 1999
Counterflow centrifugal elutriation is a noninvasive method for separating large numbers of cells on the basis of their size and mass. For mammalian cells, this method is useful for separating mixed populations of cells, in particular cells at different stages of the cell division cycle without perturbing cell metabolism or using synchronizing agents. This unit describes a method for separating 2 x 108 cells using the standard JE-6B rotor or larger numbers of cells in the JE-5.0 rotor. To verify the purity and to characterize the cell cycle positions of cells in the elutriated populations, the unit includes protocols for measuring nascent DNA synthesis by [3H]thymidine incorporation and for detecting DNA synthesis and content by propidium iodide flow cytometry alone or in combination with bromodeoxyuridine incorporation.