PNNL

Abstract

Lab-on-a-chip systems have substantially impacted the way life-sciences are explored; life on earth, however, comprises mostly of microbes, which due to their sub-micron dimensions and high mobility are more challenging to dynamically manipulate on-a-chip. To address this challenge, we developed a high resolution microfluidic system (submicrofluidics) fabricated by direct electron beam lithography that is capable of trapping single microbes and releasing them upon demand. The fabrication method enabled the integration of sub-micron indentations (400 nm) with millimetre-scale fluidic channels rapidly in a single processing step. The larger channels deliver the cell suspension and reagents, while the sub-micron indentations immobilize the cells by locally increasing the hydrodynamic resistance. By volume exclusion, single cell trapping was possible in this system without any surface treatment. By increasing the flow rate, the microbes overcome the trap barrier and pass through the narrow indentation without undergoing lysis with kinetics that depend on their size. The fabrication method and its performance are described, along with microbial characterisations using E. coli.