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<p>The mother machine (MM) is a microfluidic device designed to trap and measure single bacteria for long-term experiments, enabling the study of bacterial growth and cell cycle dynamics. In this work, the concept of a MM is adapted to fit the requirements for investigating <i>Mycobacterium smegmatis</i>, a bacterium used extensively as a model system in tuberculosis research. A MM chip was fabricated using soft lithography, and a protocol for sample preparation and filling of the side channels for mycobacteria was established. Trapped bacteria were measured using Raman spectroscopy to introduce a new analysis approach inside the MM. The subsequent data evaluation demonstrates the potential for obtaining spatially resolved chemical information on a single cell within a side channel of the MM through Raman imaging. Furthermore, the combination of Raman imaging and stable isotope labelling, as applied in this study, demonstrates the viability of bacteria inside a side channel and opens up possibilities to study cell cycles in long-term experiments.</p> Graphical abstract <p></p>

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Raman imaging for the investigation of Mycobacterium smegmatis in a mother machine

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年1月14日

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  • Ida Kalleder,
  • Eva Krois,
  • Karin Wieland,
  • Anna-Cathrine Neumann-Cip,
  • Charlott Leu,
  • Andreas Wieser,
  • Susanna Oswald,
  • Christoph Haisch

摘要

The mother machine (MM) is a microfluidic device designed to trap and measure single bacteria for long-term experiments, enabling the study of bacterial growth and cell cycle dynamics. In this work, the concept of a MM is adapted to fit the requirements for investigating Mycobacterium smegmatis, a bacterium used extensively as a model system in tuberculosis research. A MM chip was fabricated using soft lithography, and a protocol for sample preparation and filling of the side channels for mycobacteria was established. Trapped bacteria were measured using Raman spectroscopy to introduce a new analysis approach inside the MM. The subsequent data evaluation demonstrates the potential for obtaining spatially resolved chemical information on a single cell within a side channel of the MM through Raman imaging. Furthermore, the combination of Raman imaging and stable isotope labelling, as applied in this study, demonstrates the viability of bacteria inside a side channel and opens up possibilities to study cell cycles in long-term experiments.

Graphical abstract