Microfluidic guillotine for single-cell wound repair studies

Abstract

Wound repair is a key feature distinguishing living from nonliving matter. Single cells are increasingly recognized to be capable of healing wounds. The lack of reproducible, high-throughput wounding methods has hindered single-cell wound repair studies. This work describes a microfluidic guillotine for bisecting single Stentor coeruleus cells in a continuous-flow manner. Stentor is used as a model due to its robust repair capacity and the ability to perform gene knockdown in a high-throughput manner. Local cutting dynamics reveals two regimes under which cells are bisected, one at low viscous stress where cells are cut with small membrane ruptures and high viability and one at high viscous stress where cells are cut with extended membrane ruptures and decreased viability. A cutting throughput up to 64 cells per minute-more than 200 times faster than current methods-is achieved. The method allows the generation of more than 100 cells in a synchronized stage of their repair process. This capacity, combined with high-throughput gene knockdown in Stentor, enables time-course mechanistic studies impossible with current wounding methods.

Programming self-organizing multicellular structures with synthetic cell-cell signaling

“…In many cases, natural self-organized tissues have an ability to regenerate after injury (17). To test how this three-layer structure would respond to injury, we cut the structure into two fragments with a microfluidic guillotine system (18) (movie S2). Immediately after cleavage, the GFP-positive core cells were exposed to the surface, but within 24 hours, the green core cells were re-enveloped by the red layer, regenerating the spherical three-layer structure (Fig.…”

Chemical operations on a living single cell by open microfluidics for wound repair studies and organelle transport analysis

“…Wound repair of cell plasma is a vital feature distinguishing living from non-living cells 48. In some microfluidics approaches,49 the probe or key functional part was integrated into the closed microchannel and it was not possible to perform spatial movements. As a result, the cells should move toward the probe or functional part with the fluid, and thus this is not applicable to adherent cells.…”

Pulling and Pushing Stem Cells to Control Their Differentiation

“…Aided by microfluidic channels, it is possible to trap (17), capture-and-release (18), divide (19), encapsulate (20)(21)(22)(23)(24) and use for bioprinting (21) laying basis for further possibilities of genetic treatment and cell manipulation at the micro-and submicron levels. The use of single cell encapsulation will enable more possibilities of control of stem cells by inclusion of chemical factors and gradient building (25) in resulting cellbiomaterial construct.…”