Rapid mechanical deformationof cells

2PP-DLW is used to enable a microfluidic approach in which cells are mechanically deformed as they pass through a constriction smaller than the cell diameter. The compression and shear forces result in the formation of transient holes that enable the diffusion of material from the surrounding buffer into the cell. This method is employed with the aim of delivering macromolecules of interest to almost any cell type at high throughput. We are currently working on optimizing the constriction of the microfluidic chip design, considering both delivery efficiency and cell viability. The microfluidic chips were fabricated using 2PP-DLW patterning of organic-ceramic polymer compounds and consists of cell-scattering and deformation zones, where each zone contains an array of constrictions with adapted parameters. The chip was designed by arranging areas devices side by side. Experiments are  performed  to  achieve  a  better  understanding  of  the  mechanisms  of membrane disruption. The method will be used to deliver materials such as proteins and RNA to a variety of cell types including embryonic stem cells and immune cells.

Figure 5: Left: Microchannel formed in PDMS from a DLW-Ormocomp premaster with scattering (round) and a constriction (diamond) area for infection. Middle: Z-stack confocal image of a fluorescent marker inside the constriction area with varying gaps for characterization of the channel morphology. Right: Cells squeezing through the constriction/transfection area.