Improved Sedimentation Field-Flow Fractionation Separation Channel for Concentrated Cellular Elution

J Chromatogr A. 2013 Aug 9;1302:118-24. doi: 10.1016/j.chroma.2013.05.067. Epub 2013 Jun 3.

Abstract

SdFFF is now commonly used for cell sorting. Nevertheless, as with many other separation methods, SdFFF Hyperlayer elution leads (1) to sample dilution resulting in cell loss which could restrict further use; and (2) to a high output flow rate impacting detector sensitivity and selectivity. In order to limit these problems, we proposed modifications of the SdFFF separation channel consisting both in downscaling and the insertion of an outlet stream splitter. This last system corresponded to a strip which divides the flow rate output into two parts, one containing concentrated cells in a reduced volume and flow rate, the other containing the excess mobile phase useless for further cell manipulation, detection and characterization. For the first time we have shown that splitter implementation and downscaling respected channel flowing and resulted in Hyperlayer elution of around 95% of cells in less than 50% of input flow rate. Improved cell sorting was demonstrated by enrichment (∼10 times) of cancer stem cells from WiDr cells with two times less quantity of injected cells.

Keywords: Cancer stem cells; Channel downscaling; Hyperlayer elution mode; Instrument development; Outlet stream splitter; Sedimentation field flow fractionation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cell Separation / methods*
  • Fractionation, Field Flow / methods*
  • Humans
  • Neoplastic Stem Cells / cytology