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On-chip cell lysis by antibacterial non-leaching reusable quaternary ammonium monolithic column

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Abstract

Reusable antibacterial non-leaching monolithic columns polymerized in microfluidic channels designed for on-chip cell lysis applications were obtained by the photoinitiated free radical copolymerization of diallyldimethylammonium chloride (DADMAC) and ethylene glycol diacrylate (EGDA) in the presence of a porogenic solvent. The microfluidic channels were fabricated in cross-linked poly(methyl methacrylate) (X-PMMA) substrates by laser micromachining. The monolithic columns have the ability to inhibit the growth of, kill and efficiently lyse Gram-positive Micrococcus luteus (Schroeter) (ATCC 4698) and Kocuria rosea (ATCC 186), and Gram-negative bacteria Pseudomonas putida (ATCC 12633) and Escherichia coli (ATCC 35218) by mechanically shearing the bacterial membrane when forcing the cells to pass through the narrow pores of the monolithic column, and simultaneously disintegrating the cell membrane by physical contact with the antibacterial surface of the column. Cell lysis was confirmed by off-chip PCR without the need for further purification. The influence of the cross-linking monomer on bacterial growth inhibition, leaching, lysis efficiency of the monolithic column and its mechanical stability within the microfluidic channel were investigated and analyzed for three different cross-linking monomers: ethylene glycol dimethacrylate (EGDA), ethylene glycol dimethacrylate (EGDMA) and 1,6-hexanediol dimethacrylate (1,6-HDDMA). Furthermore, the bonding efficiency of two X-PMMA substrates with different cross-linking levels was studied. The monolithic columns were shown to be stable, non-leaching, and reusable for over 30 lysis cycles without significant performance degradation or DNA carryover when they were back-flushed between lysis cycles.

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Acknowledgments

We would like to express our gratitude to Grand Challenges Canada and Bigtec Labs for their financial support. We also thank Prof. Michael Palmer, Mr. Mohamed Salah, and Mr. Eric K. Brefo-Mensah, from the Department of Chemistry at the University of Waterloo, for providing access to their lab facilities and their guidance on some of the biological concepts.

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Authors and Affiliations

  1. Department of Systems Design Engineering, University of Waterloo, 200 University Ave West, Waterloo, ON, N2L 3G1, Canada

    Mohamed Aly Saad Aly & John Yeow

  2. Department of Chemistry, University of Waterloo, Waterloo, ON, Canada

    Mario Gauthier

Authors
  1. Mohamed Aly Saad Aly
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  2. Mario Gauthier
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  3. John Yeow
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Correspondence to John Yeow.

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Aly Saad Aly, M., Gauthier, M. & Yeow, J. On-chip cell lysis by antibacterial non-leaching reusable quaternary ammonium monolithic column. Biomed Microdevices 18, 2 (2016). https://doi.org/10.1007/s10544-015-0025-z

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