2D format for screening bacterial cells at the throughput of flow cytometry

Authors

  • Alexander A. Gordeev,

    1. Institute of Protein Research of the Russian Academy of Sciences, 4 Institutskaya Str., Pushchino, Moscow Region 142290, Russia; telephone: 7-496-731-8439; fax: 7-496-731-8435
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  • Timur R. Samatov,

    1. Institute of Protein Research of the Russian Academy of Sciences, 4 Institutskaya Str., Pushchino, Moscow Region 142290, Russia; telephone: 7-496-731-8439; fax: 7-496-731-8435
    Current affiliation:
    1. Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
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  • Helena V. Chetverina,

    1. Institute of Protein Research of the Russian Academy of Sciences, 4 Institutskaya Str., Pushchino, Moscow Region 142290, Russia; telephone: 7-496-731-8439; fax: 7-496-731-8435
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  • Alexander B. Chetverin

    Corresponding author
    1. Institute of Protein Research of the Russian Academy of Sciences, 4 Institutskaya Str., Pushchino, Moscow Region 142290, Russia; telephone: 7-496-731-8439; fax: 7-496-731-8435
    • Institute of Protein Research of the Russian Academy of Sciences, 4 Institutskaya Str., Pushchino, Moscow Region 142290, Russia; telephone: 7-496-731-8439; fax: 7-496-731-8435.
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Abstract

We present a method for generating gel-based unordered 2D arrays of bacterial cells of a very high density, up to 105 cells per mm2. Bacteria in a suspension are focused into a thin layer when the suspension and a dry gel matrix penetrate each other. Formation of a second gel from gel-forming components contained in the suspension results in immobilization of the cells. The immobilized cells stay alive and can repeatedly divide to produce microcolonies. The method provides for high-throughput screening and massively parallel analysis of individual cells in large populations, as well as for rapid isolation of rare clones. Biotechnol. Bioeng. 2011;108: 2682–2690. © 2011 Wiley Periodicals, Inc.

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