Agudelo Carlos G, Packirisamy Muthukumaran, Geitmann Anja
Optical Bio-Microsystem Lab, Mechanical Engineering Department, Concordia University, Montreal, QC, Canada.
Methods Mol Biol. 2014;1080:237-48. doi: 10.1007/978-1-62703-643-6_20.
A major limitation in the study of pollen tube growth has been the difficulty in providing an in vitro testing microenvironment that physically resembles the in vivo conditions. Here we describe the development of a lab-on-a-chip (LOC) for the manipulation and experimental testing of individual pollen tubes. The design was specifically tailored to pollen tubes from Camellia japonica, but it can be easily adapted for any other species. The platform is fabricated from polydimethylsiloxane (PDMS) using a silicon/SU-8 mold and makes use of microfluidics to distribute pollen grains to serially arranged microchannels. The tubes are guided into these channels where they can be tested individually. The microfluidic platform allows for specific testing of a variety of growth behavioral features as demonstrated with a simple mechanical obstacle test, and it permits the straightforward integration of further single-cell test assays.
花粉管生长研究中的一个主要限制是难以提供一个在物理上类似于体内条件的体外测试微环境。在此,我们描述了一种用于单个花粉管操作和实验测试的芯片实验室(LOC)的开发。该设计是专门为日本山茶的花粉管量身定制的,但它可以很容易地适用于任何其他物种。该平台由聚二甲基硅氧烷(PDMS)使用硅/SU-8模具制造而成,并利用微流体技术将花粉粒分布到串联排列的微通道中。花粉管被引导到这些通道中,在那里可以对它们进行单独测试。微流体平台允许对各种生长行为特征进行特定测试,如通过简单的机械障碍测试所证明的那样,并且它允许直接整合进一步的单细胞测试分析。