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Investigation of daughter cell dissection coincidence of single budding yeast cells immobilized in microfluidic traps

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Abstract

Microfluidic methodologies allow for automatic and high-throughput replicative lifespan (RLS) determination of single budding yeast cells. However, the resulted RLS is highly impacted by the robustness of experimental conditions, especially the microfluidic yeast-trapping structures, which are designed for cell retention, growth, budding, and daughter cell dissection. In this work, four microfluidic yeast-trapping structures, which were commonly used to immobilize mother cells and remove daughter cells for entire lifespan of budding yeast, were systematically investigated by means of finite element modeling (FEM). The results from this analysis led us to propose an optimized design, the yeast rotation (YRot) trap, which is a “leaky bowl”–shaped structure composed of two mirrored microcolumns facing each other. The YRot trap enables stable retention of mother cells in its “bowl” and hydrodynamic rotation of buds into its “leaky orifice” such that matured progenies can be dissected in a coincident direction. We validated the functions of the YRot trap in terms of cell rotation and daughter dissection by both FEM simulations and experiments. With the integration of denser YRot traps in microchannels, the microfluidic platform with stable single-yeast immobilization, long-term cell culturing, and coincident daughter dissection could potentially improve the robustness of experimental conditions for precise RLS determination in yeast aging studies.

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Acknowledgements

We thank Dr. Fabian Rudolf in D-BSSE, ETH Zurich, Switzerland, for providing us the yeast cell strain as a gift.

Funding

This work was supported by the National Key R&D Program of China (No. 2018YFF01012100), the National Natural Science Foundation of China (No. 61774036, 51972058, 11774051), and the Fundamental Research Funds for the Central Universities.

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Authors and Affiliations

  1. Key Laboratory of MEMS of Ministry of Education, Southeast University, Sipailou 2, Nanjing, 210096, Jiangsu, China

    Xingyu Xu, Zhen Zhu, Yingying Wang, Yangye Geng & Feng Xu

  2. School of Pharmaceutical Science and Technology, Tianjin University, Weijin Road 92, Tianjin, 300072, China

    Mario A. Marchisio

  3. School of Electronics and Information Technology, Sun Yat-Sen University, Xingang Xi Road 135, Guangzhou, 510275, Guangdong, China

    Zixin Wang

  4. CAM-SU Genomic Resource Center, Soochow University, Ren-ai Road 199, Suzhou, 215213, Jiangsu, China

    Dejing Pan

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  1. Xingyu Xu
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Xu, X., Zhu, Z., Wang, Y. et al. Investigation of daughter cell dissection coincidence of single budding yeast cells immobilized in microfluidic traps. Anal Bioanal Chem 413, 2181–2193 (2021). https://doi.org/10.1007/s00216-021-03186-x

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  • DOI: https://doi.org/10.1007/s00216-021-03186-x

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