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Microfluidic platform for integrated compartmentalization of single zoospores, germination and measurement of protrusive force generated by germ tubes
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-10-13 , DOI: 10.1039/d0lc00752h Yiling Sun 1 , Ayelen Tayagui 2 , Ashley Garrill 3 , Volker Nock 1
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-10-13 , DOI: 10.1039/d0lc00752h Yiling Sun 1 , Ayelen Tayagui 2 , Ashley Garrill 3 , Volker Nock 1
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This paper describes the design, fabrication and characterisation of a novel monolithic lab-on-a-chip (LOC) platform combining the trapping and germination of individual zoospores of the oomycete Achlya bisexualis with elastomeric micropillar-based protrusive force sensing. The oomycetes are of significant interest due to their pathogenic capabilities, which can have profound ecological and economic impacts. Zoospore encystment and germination via a germ tube play a key role in their pathogenicity. Our platform enables the study of these processes at a single cell level through hydrodynamic trapping of zoospores and their individual compartmentalization via normally closed pneumatic membrane microvalves. Valve geometry was optimized and media exchange characterized during dynamic valve operations to enhance the capture-to-growth ratio. We demonstrate germination of A. bisexualis zoospores on the platform and report three distinct germination patterns. Once germinated, germ tubes grew down growth channels towards single elastomeric micropillars. Tracking of pillar movement allowed for the measurement of microNewton range protrusive forces imparted by the tips of the germ tubes. Results indicate that the forces generated by the germ tubes are smaller than those exerted by mature hyphae. Through the use of parallel traps, channels and pillars on the same device, the platform enables high-throughput screening (HTS) of zoospores and their generation of protrusive force, an essential component of their infective capability. Due to its versatility, it will also allow for the screening of naturally bioactive compounds and the development of new biocontrol strategies for oomycetes, and morphologically similar fungal infections, as an alternative to agrochemicals.
中文翻译：
微流控平台，用于单个游动孢子的整合区室化，发芽和胚芽管产生的突出力的测量
本文介绍了一种新颖的整体式单芯片实验室（LOC）平台的设计，制造和表征，该平台结合了卵菌双性ch虫的单个游动孢子的捕获和发芽与基于弹性微柱的突出力感测相结合。卵菌的致病能力引起人们极大的兴趣，这可能会对生态和经济产生深远的影响。游动孢子的囊化和通过胚芽管的萌发在其致病性中起关键作用。我们的平台可通过游动游动孢子的流体动力捕获和它们的个别区室化，在单个细胞水平上研究这些过程常闭气动膜微阀。优化了阀门的几何形状，并在动态阀门操作过程中对介质交换进行了特征化，以提高捕获率与生长率。我们证明了双歧杆菌的发芽平台上的游动孢子并报告了三种不同的发芽模式。一旦发芽，胚芽管就会沿着生长通道向单个弹性体微柱生长。跟踪柱的运动可以测量由胚管尖端施加的微牛顿范围推力。结果表明，由胚管产生的力要比由成熟菌丝施加的力要小。通过在同一设备上使用平行阱，通道和柱子，该平台可以对游动孢子进行高通量筛选（HTS），并产生突触力，这是其感染能力的重要组成部分。由于它的多功能性，它还可以筛选天然生物活性化合物，并开发卵菌和形态相似的真菌感染的新生物防治策略，
更新日期：2020-11-03
中文翻译：
微流控平台，用于单个游动孢子的整合区室化，发芽和胚芽管产生的突出力的测量
本文介绍了一种新颖的整体式单芯片实验室（LOC）平台的设计，制造和表征，该平台结合了卵菌双性ch虫的单个游动孢子的捕获和发芽与基于弹性微柱的突出力感测相结合。卵菌的致病能力引起人们极大的兴趣，这可能会对生态和经济产生深远的影响。游动孢子的囊化和通过胚芽管的萌发在其致病性中起关键作用。我们的平台可通过游动游动孢子的流体动力捕获和它们的个别区室化，在单个细胞水平上研究这些过程常闭气动膜微阀。优化了阀门的几何形状，并在动态阀门操作过程中对介质交换进行了特征化，以提高捕获率与生长率。我们证明了双歧杆菌的发芽平台上的游动孢子并报告了三种不同的发芽模式。一旦发芽，胚芽管就会沿着生长通道向单个弹性体微柱生长。跟踪柱的运动可以测量由胚管尖端施加的微牛顿范围推力。结果表明，由胚管产生的力要比由成熟菌丝施加的力要小。通过在同一设备上使用平行阱，通道和柱子，该平台可以对游动孢子进行高通量筛选（HTS），并产生突触力，这是其感染能力的重要组成部分。由于它的多功能性，它还可以筛选天然生物活性化合物，并开发卵菌和形态相似的真菌感染的新生物防治策略，
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