Optical Bio-Microsystem Laboratory, Mechanical Engineering Department, Concordia University, Montreal, QC, Canada.
Plant J. 2013 Mar;73(6):1057-68. doi: 10.1111/tpj.12093. Epub 2013 Jan 15.
Large-scale phenotyping of tip-growing cells such as pollen tubes has hitherto been limited to very crude parameters such as germination percentage and velocity of growth. To enable efficient and high-throughput execution of more sophisticated assays, an experimental platform, the TipChip, was developed based on microfluidic and microelectromechanical systems (MEMS) technology. The device allows positioning of pollen grains or fungal spores at the entrances of serially arranged microchannels equipped with microscopic experimental set-ups. The tip-growing cells (pollen tubes, filamentous yeast or fungal hyphae) may be exposed to chemical gradients, microstructural features, integrated biosensors or directional triggers within the modular microchannels. The device is compatible with Nomarski optics and fluorescence microscopy. Using this platform, we were able to answer several outstanding questions on pollen tube growth. We established that, unlike root hairs and fungal hyphae, pollen tubes do not have a directional memory. Furthermore, pollen tubes were found to be able to elongate in air, raising the question of how and where water is taken up by the cell. The platform opens new avenues for more efficient experimentation and large-scale phenotyping of tip-growing cells under precisely controlled, reproducible conditions.
大规模表型分析如花粉管这样的顶端生长细胞,迄今为止一直局限于非常粗略的参数,如发芽率和生长速度。为了能够高效且高通量地执行更复杂的检测,我们基于微流控和微机电系统(MEMS)技术开发了一个实验平台,称为 TipChip。该设备允许将花粉粒或真菌孢子定位在依次排列的微通道的入口处,这些微通道配备有微观实验装置。顶端生长细胞(花粉管、丝状酵母或真菌菌丝)可以暴露于化学梯度、微结构特征、集成生物传感器或模块化微通道内的定向触发器中。该设备与诺马斯基光学和荧光显微镜兼容。使用该平台,我们能够回答关于花粉管生长的几个悬而未决的问题。我们确定,与根毛和真菌菌丝不同,花粉管没有方向记忆。此外,发现花粉管能够在空气中伸长,这就提出了一个问题,即细胞如何以及在何处吸收水分。该平台为在精确控制、可重复的条件下更有效地进行顶端生长细胞的实验和大规模表型分析开辟了新途径。