Leung Jacob C K, Hilliker Arthur J, Rezai Pouya
Department of Mechanical Engineering, York University, BCEE 433B, 4700 Keele St, Toronto, ON M3J 1P3, Canada.
Lab Chip. 2016 Feb 21;16(4):709-19. doi: 10.1039/c5lc01517k. Epub 2016 Jan 15.
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.
使用黑腹果蝇(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。在化学掺杂的琼脂平板上产卵是用于定量评估果蝇生物学适应性和行为反应的重要读数指标。由于琼脂的手动化学掺杂,当前基于产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,我们开发了一种新型的琼脂 - 聚二甲基硅氧烷(PDMS)混合微流控装置,用于对自由飞行的成年果蝇进行单浓度和多浓度化学给药以及芯片上的产卵筛选。为实现这一目标,我们设计了一种新技术,以冰作为牺牲层将琼脂与PDMS通道集成。随后,我们使用不同浓度的锌和乙酸对成年黑腹果蝇进行了单化学毒性和多选择化学偏好试验。我们的装置使我们能够:1)证明果蝇能够在PDMS - 琼脂微流控装置上感知不同化学物质的浓度,这在确定产卵位点选择中起重要作用;2)研究单浓度和多浓度化学环境之间的产卵偏好是否不同。该装置可用于研究果蝇和其他产卵昆虫的基础和应用生物学问题。它在设计上还可以扩展,以开发未来复杂且动态的化学给药和高通量筛选平台,而这是现有产卵筛选技术难以实现的。