An Inert Continuous Microreactor for the Isolation and Analysis of a Single Microbial Cell

Micromachines

Katrin Rosenthal³  Floris Falke²  Oliver Frick¹  Christian Dusny¹  Andreas Schmid¹ 

[1] Department Solar Materials, Helmholtz Centre for Environmental Research (UFZ), Permoser Str. 15, D-04318 Leipzig, Germany;LioniX B.V., Hengelosestraat 500, 7521 AN Enschede, The Netherlands;Laboratory of Chemical Biotechnology, Department of Biochemical & Chemical Engineering, TU Dortmund University, Emil-Figge-Str. 66, D-44227 Dortmund, Germany;

关键词: lab-on-a-chip;    negative dielectrophoresis;    direct fusion bonding;    single-cell analysis;    Corynebacterium glutamicum;   

DOI  :  10.3390/mi6121459

来源: mdpi

【 摘 要 】

Studying biological phenomena of individual cells is enabled by matching the scales of microbes and cultivation devices. We present a versatile, chemically inert microfluidic lab-on-a-chip (LOC) device for biological and chemical analyses of isolated microorganisms. It is based on the Envirostat concept and guarantees constant environmental conditions. A new manufacturing process for direct fusion bonding chips with functional microelectrodes for selective and gentle cell manipulation via negative dielectrophoresis (nDEP) was generated. The resulting LOC system offered a defined surface chemistry and exceptional operational stability, maintaining its structural integrity even after harsh chemical treatment. The microelectrode structures remained fully functional after thermal bonding and were proven to be efficient for single-cell trapping via nDEP. The microfluidic network consisted solely of glass, which led to enhanced chip reusability and minimized interaction of the material with chemical and biological compounds. We validated the LOC for single-cell studies with the amino acid secreting bacterium Corynebacterium glutamicum. Intracellular l-lysine production dynamics of individual bacteria were monitored based on a genetically encoded fluorescent nanosensor. The results demonstrate the applicability of the presented LOC for pioneering chemical and biological studies, where robustness and chemically inert surfaces are crucial parameters for approaching fundamental biological questions at a single-cell level.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.