title: Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics.
abstract: 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.
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Tao
authors.initials: T
authors.lastname: Geng
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Erin L
authors.initials: EL
authors.lastname: Bredeweg
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Szymanski
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Bingwen
authors.initials: B
authors.lastname: Liu
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Baker
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Galya
authors.initials: G
authors.lastname: Orr
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Evans
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Ryan T
authors.initials: RT
authors.lastname: Kelly
mesh_terms: D002244:Carbon / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D025301:Hyphae / Q000502:physiology; D046228:Microarray Analysis; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D009024:Morphogenesis; D009492:Neurospora crassa / Q000254:growth & development* / Q000378:metabolism
chemical_list: D002244:Carbon