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Digital microfluidic immunocytochemistry in single cells

Author

Listed:
  • Alphonsus H. C. Ng

    (Institute of Biomaterials and Biomedical Engineering, University of Toronto
    Donnelly Centre for Cellular and Biomolecular Research)

  • M. Dean Chamberlain

    (Institute of Biomaterials and Biomedical Engineering, University of Toronto
    Donnelly Centre for Cellular and Biomolecular Research
    Department of Chemical Engineering and Applied Chemistry)

  • Haozhong Situ

    (Institute of Biomaterials and Biomedical Engineering, University of Toronto
    Donnelly Centre for Cellular and Biomolecular Research)

  • Victor Lee

    (Donnelly Centre for Cellular and Biomolecular Research)

  • Aaron R. Wheeler

    (Institute of Biomaterials and Biomedical Engineering, University of Toronto
    Donnelly Centre for Cellular and Biomolecular Research
    University of Toronto)

Abstract

We report a new technique called Digital microfluidic Immunocytochemistry in Single Cells (DISC). DISC automates protocols for cell culture, stimulation and immunocytochemistry, enabling the interrogation of protein phosphorylation on pulsing with stimulus for as little as 3 s. DISC was used to probe the phosphorylation states of platelet-derived growth factor receptor (PDGFR) and the downstream signalling protein, Akt, to evaluate concentration- and time-dependent effects of stimulation. The high time resolution of the technique allowed for surprising new observations—for example, a 10 s pulse stimulus of a low concentration of PDGF is sufficient to cause >30% of adherent fibroblasts to commit to Akt activation. With the ability to quantitatively probe signalling events with high time resolution at the single-cell level, we propose that DISC may be an important new technique for a wide range of applications, especially for screening signalling responses of a heterogeneous cell population.

Suggested Citation

  • Alphonsus H. C. Ng & M. Dean Chamberlain & Haozhong Situ & Victor Lee & Aaron R. Wheeler, 2015. "Digital microfluidic immunocytochemistry in single cells," Nature Communications, Nature, vol. 6(1), pages 1-12, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8513
    DOI: 10.1038/ncomms8513
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