Lab-on-a-chip for cell perturbation, lysis, and efficient separation of sub-cellular components in a continuous flow mode

• We developed a novel microfluidic device for the preparation of cell samples.
• Cells are on-chip perturbed and permeabilized followed by separation of organelles.
• An efficient method is presented for rapid separation of sub-cellular components.
• The chip comprises two adjacent temperature regions which are thermally decoupled.
• An integrated chip-bioreactor setup allows dynamic experiments in continuous flow.

A novel lab-on-a-chip is presented for the high-throughput preparation of mammalian cell samples for metabolic studies associated with cell compartmentation. The chip is directly connected to an overpressurized bioreactor, for a continuous harvest and manipulation of cells during the cultivation process. The proposed on-chip preparation technique allows the rapid cell permeabilization and separation of sub-cellular components as cytoplasm and mitochondria, both primarily responsible for cell metabolism. The microfluidic chip comprises two temperature zones (37 °C to ensure physiological conditions and 4 °C for cell quenching), which are thermally decoupled. Cells are manipulated in a continuous flow process in five connected functional modules. The key modules of the integrated chip are a rapid split-and-recombine micromixer for biological system perturbation and chemical lysis, a chaotic advection “stirred” incubation channel for reduced residence time distribution, and a spiral channel for efficient particle separation and media exchange. The bioreactor-chip system was characterized with Chinese hamster ovary cells. It exhibits a high efficiency in the lysis process and separation of sub-cellular components.