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首页> 外文期刊>Biochemistry >A Microfluidic-Based Microscopy Platform for Continuous Interrogation of Trypanosoma brucei during Environmental Perturbation
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A Microfluidic-Based Microscopy Platform for Continuous Interrogation of Trypanosoma brucei during Environmental Perturbation

机译：基于微流体的显微镜平台，用于在环境扰动期间连续询问锥虫瘤斑瘤

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摘要

The African trypanosome, Trypanosoma brucei, is the causative agent of human African trypanosomiasis (HAT). African trypanosomes are extracellular parasites that possess a single flagellum that imparts a high degree of motility to the microorganisms. In addition, African trypanosomes show significant metabolic and structural adaptation to environmental conditions. Analysis of the ways that environmental cues affect these organisms generally requires rapid perfusion experiments in combination with single-cell imaging, which are difficult to apply under conditions of rapid motion. Microfluidic devices have been used previously as a strategy for trapping small motile cells in a variety of organisms, including trypanosomes; however, in the past, such devices required individual fabrication in a cleanroom, limiting their application. Here we demonstrate that a commercial microfluidic device, typically used for bacterial trapping, can trap bloodstream and procyclic form trypanosomes, allowing for rapid buffer exchange via perfusion. As a result, time-lapse single-cell microscopy images of these highly motile parasites were acquired during environmental variations. Using these devices, we have been able to perform and analyze perfusion-based single-cell tracking experiments of the responses of the parasite to changes in glucose availability, which is a major step in resolving the mechanisms of adaptation of kinetoplasts to their individual biological niches; we demonstrate utility of this tool for making measurements of procyclic form trypanosome intracellular glucose levels as a function of changes in extracellular glucose concentrations. These experiments demonstrate that cytosolic glucose equilibrates with external conditions as fast as, or faster than, the rate of solution exchange in the instrument.
机译：非洲锥虫葡萄质瘤斑瘤布氏蛋白酶，是人类非洲锥虫病（帽子）的致病因子。非洲锥虫是细胞外寄生虫，其具有单一的鞭毛，其赋予微生物高度的运动性。此外，非洲锥虫表现出对环境条件的显着代谢和结构调整。对环境提示影响这些生物的方式的分析通常需要快速灌注实验与单细胞成像组合，这很难在快速运动的条件下施加。以前使用的微流体装置作为捕获各种生物体中的小运动细胞的策略，包括锥虫;然而，在过去，这种装置需要在洁净室中的个体制造，限制它们的应用。在这里，我们证明通常用于细菌捕获的商业微流体装置可以捕获血液和药物形状的锥虫，允许通过灌注快速交换。结果，在环境变化期间获得了这些高运动寄生虫的时间流逝单细胞显微镜图像。使用这些设备，我们能够对寄生虫的反应进行灌注的单细胞跟踪实验，以改变葡萄糖可用性，这是解决动力骨架适应其个体生物核桃的机制的重要步骤;我们展示了该工具的效用，用于测量胰腈形成锥虫细胞内葡萄糖水平，作为细胞外葡萄糖浓度的变化。这些实验表明，胞质葡萄糖与仪器中的溶液交换速率快速或更快的外部条件平衡。

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  • 来源
    《Biochemistry》 |2019年第7期|共8页
  • 作者

    Voyton Charles M.; Choi Jongsu; Qiu Yijian; Morris Meredith T.; Ackroyd P. Christine; Morris James C.; Christensen Kenneth A.;

  • 作者单位

    Clemson Univ Dept Chem Clemson SC 29634 USA;

    Brigham Young Univ Dept Chem &

    Biochem Provo UT 84602 USA;

    Clemson Univ Dept Genet &

    Biochem Eukaryot Pathogens Innovat Ctr Clemson SC 29634 USA;

    Clemson Univ Dept Genet &

    Biochem Eukaryot Pathogens Innovat Ctr Clemson SC 29634 USA;

    Brigham Young Univ Dept Chem &

    Biochem Provo UT 84602 USA;

    Clemson Univ Dept Genet &

    Biochem Eukaryot Pathogens Innovat Ctr Clemson SC 29634 USA;

    Clemson Univ Dept Chem Clemson SC 29634 USA;

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  • 正文语种 eng
  • 中图分类 生物化学;
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