Hydrodynamic dissection of <i>Stentor coeruleus</i> in a microfluidic cross junction.

Abstract

<i>Stentor coeruleus</i>, a single-cell ciliated protozoan, is a model organism for wound healing and regeneration studies. Despite <i>Stentor</i>'s large size (up to 2 mm in extended state), microdissection of <i>Stentor</i> remains challenging. In this work, we describe a hydrodynamic cell splitter, consisting of a microfluidic cross junction, capable of splitting <i>Stentor</i> 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 <i>Stentor</i> 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 <i>Stentor</i>.