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An integrated hybrid microfluidic device for oviposition-based chemical screening of adult Drosophila melanogaster

机译:一种集成的混合微流控设备,用于基于成虫果蝇产卵的化学筛选

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Chemical screening using Drosophila melanogaster (the fruit fly) is vital in drug discovery, agricultural, and toxicological applications. Oviposition (egg laying) on chemically-doped agar plates is an important read-out metric used to quantitatively assess the biological fitness and behavioral responses of Drosophila. Current oviposition-based chemical screening studies are inaccurate, labor-intensive, time-consuming, and inflexible due to the manual chemical doping of agar. In this paper, we have developed a novel hybrid agar-polydimethylsiloxane (PDMS) microfluidic device for single-and multi-concentration chemical dosing and on-chip oviposition screening of free-flying adult stage Drosophila. To achieve this, we have devised a novel technique to integrate agar with PDMS channels using ice as a sacrificial layer. Subsequently, we have conducted single-chemical toxicity and multiple choice chemical preference assays on adult Drosophila melanogaster using zinc and acetic acid at various concentrations. Our device has enabled us to 1) demonstrate that Drosophila is capable of sensing the concentration of different chemicals on a PDMS-agar microfluidic device, which plays significant roles in determining oviposition site selection and 2) investigate whether oviposition preference differs between single-and multi-concentration chemical environments. This device may be used to study fundamental and applied biological questions in Drosophila and other egg laying insects. It can also be extended in design to develop sophisticated and dynamic chemical dosing and high-throughput screening platforms in the future that are not easily achievable with the existing oviposition screening techniques.
机译:使用果蝇(果蝇)进行化学筛选在药物发现,农业和毒理学应用中至关重要。在化学掺杂的琼脂板上产卵(卵产)是一种重要的读出指标,用于定量评估果蝇的生物学适应性和行为反应。由于琼脂的人工化学掺杂,目前基于排卵的化学筛选研究不准确,劳动强度大,耗时且不灵活。在本文中,我们开发了一种新颖的混合琼脂-聚二甲基硅氧烷(PDMS)微流控装置,用于单次飞行和多倍浓度化学计量以及片上产卵果蝇的果蝇产卵。为实现此目的,我们设计了一种新技术,可将琼脂与PDMS通道整合在一起,并使用冰作为牺牲层。随后,我们使用不同浓度的锌和乙酸对成年果蝇进行了单化学毒性和多选化学偏好分析。我们的设备使我们能够:1)证明果蝇能够检测PDMS-琼脂微流控设备上不同化学物质的浓度,这在确定产卵位点选择中起着重要作用; 2)研究单卵胎和多胎卵子的产卵偏好是否不同浓度化学环境。该设备可用于研究果蝇和其他产卵昆虫的基本和应用生物学问题。它还可以在设计上进行扩展,以开发将来无法使用现有的排卵筛查技术轻松实现的复杂,动态的化学计量和高通量筛查平台。

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