A microfluidic single-cell array for in situ laminar-flow-based comparative culturing of budding yeast cells

Zhu, Zhen; Wang, Yingying; Peng, Ruobo; Chen, Pan; Geng, Yangye; He, Bai-Liang; Ouyang, Shuiping; Zheng, Ke; Fan, Yimin; Pan, Dandan; Jin, Nan; Rudolf, Fabian; Hierlemann, Andreas

doi:10.1016/j.talanta.2021.122401

Cited by 5 publications

(3 citation statements)

References 49 publications

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“…We developed a microfluidic device with an array of single-cell traps for reliable immobilization of single yeast cells and long-term cell culturing under precisely controlled medium perfusion. The device also features rapid medium switching and microscopic monitoring of the in-situ XCM behavior at single cell level [ 30 ] (Fig. 2 A).…”

Section: Resultsmentioning

confidence: 99%

Protein acetylation regulates xylose metabolism during adaptation of Saccharomyces cerevisiae

Tan

Wang

et al. 2021

Biotechnol Biofuels

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Background As the second most abundant polysaccharide in nature, hemicellulose can be degraded to xylose as the feedstock for bioconversion to fuels and chemicals. To enhance xylose conversion, the engineered Saccharomyces cerevisiae with xylose metabolic pathway is usually adapted with xylose as the carbon source in the laboratory. However, the mechanism under the adaptation phenomena of the engineered strain is still unclear. Results In this study, xylose-utilizing S. cerevisiae was constructed and used for the adaptation study. It was found that xylose consumption rate increased 1.24-fold in the second incubation of the yYST12 strain in synthetic complete-xylose medium compared with the first incubation. The study figured out that it was observed at the single-cell level that the stagnation time for xylose utilization was reduced after adaptation with xylose medium in the microfluidic device. Such transient memory of xylose metabolism after adaptation with xylose medium, named “xylose consumption memory”, was observed in the strains with both xylose isomerase pathway and xylose reductase and xylitol dehydrogenase pathways. In further, the proteomic acetylation of the strains before and after adaptation was investigated, and it was revealed that H4K5 was one of the most differential acetylation sites related to xylose consumption memory of engineered S. cerevisiae. We tested 8 genes encoding acetylase or deacetylase, and it was found that the knockout of the GCN5 and HPA2 encoding acetylases enhanced the xylose consumption memory. Conclusions The behavior of xylose consumption memory in engineered S. cerevisiae can be successfully induced with xylose in the adaptation. H4K5Ac and two genes of GCN5 and HPA2 are related to xylose consumption memory of engineered S. cerevisiae during adaptation. This study provides valuable insights into the xylose adaptation of engineered S. cerevisiae.

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Section: Resultsmentioning

confidence: 99%

Protein acetylation regulates xylose metabolism during adaptation of Saccharomyces cerevisiae

Tan

Wang

et al. 2021

Biotechnol Biofuels

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“…The generation of well-controlled spatial and temporal concentration distributions of chemical substances plays a vital role in the field of chemical and biological sciences, with diverse applications ranging from synthesis, protein crystallization, − cell culture, − drug screening, , to the pharmaceutical industry . Traditional methods, such as pipettes and multiwell plates, often struggle to generate gradients accurately and stably while using minimal amounts of reactants.…”

Section: Introductionmentioning

confidence: 99%

Generation of Multiple Concentration Gradients Using a Two-Dimensional Pyramid Array

Qi,

Zhou,

et al. 2023

Anal. Chem.

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Concentration heterogeneity of diffusible reactants is a prevalent phenomenon in biochemical processes, requiring the generation of concentration gradients for the relevant experiments. In this study, we present a high-density pyramid array microfluidic network for the effective and precise generation of multiple concentration gradients. The complex gradient distribution in the 2D array can be adaptively adjusted by modulating the reactant velocities and concentrations at the inlets. In addition, the unique design of each reaction chamber and mixing block in the array ensures uniform concentrations within each chamber during dynamic changes, enabling large-scale reactions with low reactant volumes. Through detailed numerical simulation of mass transport within the complex microchannel networks, the proposed method allows researchers to determine the desired number of reaction chambers within a given concentration range based on experimental requirements and to quickly obtain the operating conditions with the help of machine learning-based prediction. The effectiveness in generating a multiple concentration gradient environment was further demonstrated by concentration-dependent calcium carbonate crystallization experiments. This device provides a highly efficient mixing and adaptable concentration platform that is well suited for high-throughput and multiplexed reactions.

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“…In contact-based approaches, cells are captured by means of hydrodynamic forces and require the use of physical obstacles within the trapping region. These obstacles include features such as microchambers, 35,36 microvalves, 37,38 as well as pillars, 39,40 walls 41 or pores 42 arranged in various arrays and patterns. In general, these methods are non-flow through methods, meaning that the cells will remain captured within the trapping region for long term observation or analysis.…”

mentioning

confidence: 99%

Acoustofluidic cell micro-dispenser for single cell trajectory control

et al. 2022

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A microfluidic single-cell array for in situ laminar-flow-based comparative culturing of budding yeast cells

Cited by 5 publications

References 49 publications

Protein acetylation regulates xylose metabolism during adaptation of Saccharomyces cerevisiae

Protein acetylation regulates xylose metabolism during adaptation of Saccharomyces cerevisiae

Generation of Multiple Concentration Gradients Using a Two-Dimensional Pyramid Array

Acoustofluidic cell micro-dispenser for single cell trajectory control

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