Automated leukocyte processing by microfluidic deterministic lateral displacement

Curt I. Civin, Tony Ward, Alison M. Skelley, Khushroo Gandhi, Zendra Peilun Lee, Christopher R. Dosier, Joseph L. D'Silva, Yu Chen, Min Jung Kim, James Moynihan, Xiaochun Chen, Lee Aurich, Sergei Gulnik, George C. Brittain, Diether J. Recktenwald, Robert H. Austin, James C. Sturm

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

We previously developed a Deterministic Lateral Displacement (DLD) microfluidic method in silicon to separate cells of various sizes from blood (Davis et al., Proc Natl Acad Sci 2006;103:14779-14784; Huang et al., Science 2004;304:987-990). Here, we present the reduction-to-practice of this technology with a commercially produced, high precision plastic microfluidic chip-based device designed for automated preparation of human leukocytes (white blood cells; WBCs) for flow cytometry, without centrifugation or manual handling of samples. After a human blood sample was incubated with fluorochrome-conjugated monoclonal antibodies (mAbs), the mixture was input to a DLD microfluidic chip (microchip) where it was driven through a micropost array designed to deflect WBCs via DLD on the basis of cell size from the Input flow stream into a buffer stream, thus separating WBCs and any larger cells from smaller cells and particles and washing them simultaneously. We developed a microfluidic cell processing protocol that recovered 88% (average) of input WBCs and removed 99.985% (average) of Input erythrocytes (red blood cells) and >99% of unbound mAb in 18 min (average). Flow cytometric evaluation of the microchip Product, with no further processing, lysis or centrifugation, revealed excellent forward and side light scattering and fluorescence characteristics of immunolabeled WBCs. These results indicate that cost-effective plastic DLD microchips can speed and automate leukocyte processing for high quality flow cytometry analysis, and suggest their utility for multiple other research and clinical applications involving enrichment or depletion of common or rare cell types from blood or tissue samples.

Original languageEnglish (US)
Pages (from-to)1073-1083
Number of pages11
JournalCytometry Part A
Volume89
Issue number12
DOIs
StatePublished - Dec 1 2016

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

Keywords

  • blood
  • cell processing
  • cell sorting
  • deterministic lateral displacement
  • leukocytes
  • microfluidic
  • red blood cells
  • sample preparation
  • white blood cells

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  • Cite this

    Civin, C. I., Ward, T., Skelley, A. M., Gandhi, K., Peilun Lee, Z., Dosier, C. R., D'Silva, J. L., Chen, Y., Kim, M. J., Moynihan, J., Chen, X., Aurich, L., Gulnik, S., Brittain, G. C., Recktenwald, D. J., Austin, R. H., & Sturm, J. C. (2016). Automated leukocyte processing by microfluidic deterministic lateral displacement. Cytometry Part A, 89(12), 1073-1083. https://doi.org/10.1002/cyto.a.23019