A Microfluidic Bioreactor Based on Hydrogel-Entrapped E. coli: Cell Viability, Lysis, and Intracellular Enzyme Reactions

Abstract

Viable E. coli cells were entrapped in hydrogel micropatches photopolymerized within microfluidic systems. The microfluidic channels and the micropatches have sizes on the order of 100-500 mum. Small molecules, such as dyes and surfactants, present in the solution surrounding the hydrogel, are able to diffuse into the gel and encounter the cells, but the cells are sufficiently large to be retained. For example, sodium dodecyl sulfate is a lysis agent that is able to penetrate the hydrogel and disrupt the cellular membrane. Entrapment of viable cells within hydrogels, followed by lysis, could provide a convenient means for preparing biocatalysts without the need for enzyme extraction and purification. Hydrogel-immobilized cells are able to carry out chemical reactions within microfluidic channels. Specifically, a nonfluorescent dye, BCECF-AM, is able to penetrate both the hydrogel and the bacterial membrane and be converted into a fluorescent form (BCECF) by the interior cellular machinery. These results suggest that cells immobilized within microfluidic channels can act as sensors for small molecules and as bioreactors for carrying out reactions.