Microfluidic System for Cell Mixing and Particle Focusing Using Dean Flow Fractionation

Abstract

Recent developments in the field of additive manufacturing processes have led to tremendous technological progress and opened directions for the field of microfluidics. For instance, new flexible materials for 3D printing allow the substitution of polydimethylsiloxane (PDMS) in microfluidic prototype development. Three-dimensional-printed microfluidic components open new horizons, in particular for the automated handling of biological cells (e.g., eukaryotic cells or bacteria). Here, we demonstrate how passive mixing and passive separation processes of biological cells can be realized using 3D printing concepts for rapid prototyping. This technique facilitates low-cost experimental setups that are easy to modify and adopt for specific detection and diagnostic purposes. In particular, printing technologies based on fused deposition modeling and stereolithography are used and their realization is discussed. Additive technologies enable the fabrication of multiplication mixers, which overcome shortcomings of current pillar or curve-based techniques and enable efficient mixing, also of biological cells without affecting viability. Using standard microfluidic components and state-of-the art 3D printing technologies, we realize a separation system based on Dean flow fragmentation without the use of PDMS. In particular, we describe the use of a 3D-printed helix for winding a capillary for particle flow and a new chip design for particle separation at the outlet. We demonstrate the functionality of the system by successful isolation of ~12 µm-sized particles from a particle mixture containing large (~12 µm, typical size of eukaryotic cells) and small (~2 µm, typical size of bacteria or small yeasts) particles. Using this setup to separate eukaryotic cells from bacteria, we could prove that cell viability is not affected by passage through the microfluidic systems.