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

摘要

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 subcellular 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.