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Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics

机译:分区微通道阵列用于单细胞极化生长和动力学的高通量分析

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摘要

Interrogating polarized growth is technologically challenging due to extensive cellular branching and uncontrollable environmental conditions in conventional assays. Here we present a robust and high-performance microfluidic system that enables observations of polarized growth with enhanced temporal and spatial control over prolonged periods. The system has built-in tunability and versatility to accommodate a variety of scientific applications requiring precisely controlled environments. Using the model filamentous fungus, Neurospora crassa, our microfluidic system enabled direct visualization and analysis of cellular heterogeneity in a clonal fungal cell population, nuclear distribution and dynamics at the subhyphal level, and quantitative dynamics of gene expression with single hyphal compartment resolution in response to carbon source starvation and exchange. Although the microfluidic device is demonstrated on filamentous fungi, the technology is immediately extensible to a wide array of other biosystems that exhibit similar polarized cell growth, with applications ranging from bioenergy production to human health.
机译:由于常规检测中广泛的细胞分支和不可控制的环境条件,询问极化生长在技术上具有挑战性。在这里,我们介绍了一个强大而高性能的微流体系统,该系统能够观察极化增长,并在较长时期内增强时空控制。该系统具有内置的可调性和多功能性,可以适应需要精确控制环境的各种科学应用。使用模型丝状真菌Neurospora crassa,我们的微流体系统能够直接可视化和分析克隆真菌细胞群体中的细胞异质性,亚菌丝水平的核分布和动力学,以及响应于单个菌丝区室分辨率的基因表达的定量动力学碳源的饥饿与交换。尽管微流控设备已在丝状真菌上进行了展示,但该技术可立即扩展到其他具有相似极化细胞生长功能的其他生物系统,其应用范围从生物能源生产到人类健康。

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