Department of Mechanical Engineering, Tufts University, Medford, United States.
Department of Biology, Wake Forest University, Winston-Salem, United States.
Elife. 2023 Jul 24;12:e85348. doi: 10.7554/eLife.85348.
Microorganism sensing of and responding to ambient chemical gradients regulates a myriad of microbial processes that are fundamental to ecosystem function and human health and disease. The development of efficient, high-throughput screening tools for microbial chemotaxis is essential to disentangling the roles of diverse chemical compounds and concentrations that control cell nutrient uptake, chemorepulsion from toxins, and microbial pathogenesis. Here, we present a novel microfluidic multiplexed chemotaxis device (MCD) which uses serial dilution to simultaneously perform six parallel bacterial chemotaxis assays that span five orders of magnitude in chemostimulant concentration on a single chip. We first validated the dilution and gradient generation performance of the MCD, and then compared the measured chemotactic response of an established bacterial chemotaxis system () to a standard microfluidic assay. Next, the MCD's versatility was assessed by quantifying the chemotactic responses of different bacteria () to different chemoattractants and chemorepellents. The MCD vastly accelerates the chemotactic screening process, which is critical to deciphering the complex sea of chemical stimuli underlying microbial responses.
微生物对环境化学梯度的感知和响应调节着众多微生物过程,这些过程对生态系统功能和人类健康与疾病至关重要。开发高效、高通量的微生物趋化性筛选工具对于阐明控制细胞营养吸收、毒素化学斥力以及微生物发病机制的各种化学化合物和浓度的作用至关重要。在这里,我们提出了一种新颖的微流控多路趋化性装置(MCD),该装置使用连续稀释来同时在单个芯片上进行六个平行的细菌趋化性测定,涵盖了化学刺激物浓度的五个数量级。我们首先验证了 MCD 的稀释和梯度生成性能,然后比较了已建立的细菌趋化性系统()对标准微流控测定的测量趋化响应。接下来,通过量化不同细菌()对不同趋化剂和化学抑制剂的趋化反应来评估 MCD 的多功能性。MCD 大大加速了趋化性筛选过程,这对于破译微生物反应背后复杂的化学刺激海洋至关重要。