Department of Mechanical Engineering, Stanford University, Stanford, California, USA.
Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA.
Lab Chip. 2017 Mar 14;17(6):1116-1127. doi: 10.1039/c6lc01165a.
New tools for applying force to animals, tissues, and cells are critically needed in order to advance the field of mechanobiology, as few existing tools enable simultaneous imaging of tissue and cell deformation as well as cellular activity in live animals. Here, we introduce a novel microfluidic device that enables high-resolution optical imaging of cellular deformations and activity while applying precise mechanical stimuli to the surface of the worm's cuticle with a pneumatic pressure reservoir. To evaluate device performance, we compared analytical and numerical simulations conducted during the design process to empirical measurements made with fabricated devices. Leveraging the well-characterized touch receptor neurons (TRNs) with an optogenetic calcium indicator as a model mechanoreceptor neuron, we established that individual neurons can be stimulated and that the device can effectively deliver steps as well as more complex stimulus patterns. This microfluidic device is therefore a valuable platform for investigating the mechanobiology of living animals and their mechanosensitive neurons.
为了推动机械生物学领域的发展,迫切需要新的工具来对动物、组织和细胞施加力,因为很少有现有的工具能够同时对组织和细胞变形以及活体动物中的细胞活性进行成像。在这里,我们介绍了一种新颖的微流控装置,该装置能够在向虫体角质层表面施加气动压力储器的精确机械刺激的同时,实现对细胞变形和活性的高分辨率光学成像。为了评估设备性能,我们将设计过程中的分析和数值模拟与使用制造设备进行的经验测量进行了比较。利用作为机械感受器神经元模型的带有光遗传学钙指示剂的良好表征的触须感觉神经元 (TRN),我们确定可以刺激单个神经元,并且该设备可以有效地传递阶跃以及更复杂的刺激模式。因此,这种微流控装置是研究活体动物及其机械敏感神经元的机械生物学的有价值的平台。