Hydrodynamic dissection of Stentor coeruleus in a microfluidic cross junction
Hydrodynamic dissection of Stentor coeruleus in a microfluidic cross junction
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DOI：
10.1039/d2lc00527a
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发表时间：
2022-08-08
期刊：
影响因子：
6.1
通讯作者：
Tang，Sindy K. Y.
中科院分区：
文献类型：
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作者：
Paul，Rajorshi;Zhang，Kevin S.;Tang，Sindy K. Y.
蓝斑圆胸蛛是一种单细胞纤毛原生动物，是研究创伤愈合和再生的模式生物。尽管Stentor的尺寸较大（伸展状态下可达2 mm），但Stentor的显微切割仍然具有挑战性。在这项工作中，我们描述了一种流体动力学细胞分裂器，由一个微流体交叉路口，能够分裂Stentor细胞在一个非接触的方式在一个高通量的500个细胞每分钟连续操作下。在交叉连接处的流场中引入不对称性导致细胞的不对称分裂，从而产生小至原始细胞尺寸的1.85倍的细胞碎片。细胞碎片活力的表征显示，随着碎片大小的减小和施加在碎片上的流体动力学应力的程度的增加，5天存活率降低。我们的研究结果表明，细胞碎片的大小和组成，以及机械应力，发挥重要作用，在长期修复的Stentor细胞，并值得进一步调查。然而，流体动力学分裂器可用于研究细胞分裂后立即发生的现象，例如在Stentor中发生在100-1000秒量级的质膜中伤口的闭合。
Stentor coeruleus, a single-cell ciliated protozoan, is a model organism for wound healing and regeneration studies. Despite Stentor's large size (up to 2 mm in extended state), microdissection of Stentor remains challenging. In this work, we describe a hydrodynamic cell splitter, consisting of a microfluidic cross junction, capable of splitting Stentor cells in a non-contact manner at a high throughput of ∼500 cells per minute under continuous operation. Introduction of asymmetry in the flow field at the cross junction leads to asymmetric splitting of the cells to generate cell fragments as small as ∼8.5 times the original cell size. Characterization of cell fragment viability shows reduced 5-day survival as fragment size decreases and as the extent of hydrodynamic stress imposed on the fragments increases. Our results suggest that cell fragment size and composition, as well as mechanical stress, play important roles in the long-term repair of Stentor cells and warrant further investigations. Nevertheless, the hydrodynamic splitter can be useful for studying phenomena immediately after cell splitting, such as the closure of wounds in the plasma membrane which occurs on the order of 100–1000 seconds in Stentor.