This paper reports the development, modeling, and testing of an original microfluidic chip capable of generating both time-evolving and spatially varying gradients in standard Petri dishes. It consists of three sets of five independently controlled parallel channels, and its architecture allows the generation of complex gradient profiles that can be flexibly positioned and dynamically altered in an open cell-chamber environment. A detailed fabrication protocol for the production of these chips using multilayer soft lithography is reported. A comprehensive computational model is also presented based on COMSOL Multiphysics software that includes both diffusion and advection of the fluid as it exits the microchannels. The results of the simulation are successfully applied to model single-channel experiments. The chip is then tested in multi-channel mode, and its ability to produce complex spatially varied concentration profiles is demonstrated. The achievement of steady state of the gradient profile in less than 5 min also allows for the dynamic variation of the profile. Finally, we apply the present chip architecture to investigate the migration of mouse neutrophils in an Interleukin-8 gradient. We report quantitatively on cell migration driven by Interleukin-8 gradient and provide migration speed distribution.

Microfluidic chip for spatially and temporally controlled biochemical gradient generation in standard cell-culture Petri dishes

SAHAI, RANJANA;BELTRAM, Fabio
2011

Abstract

This paper reports the development, modeling, and testing of an original microfluidic chip capable of generating both time-evolving and spatially varying gradients in standard Petri dishes. It consists of three sets of five independently controlled parallel channels, and its architecture allows the generation of complex gradient profiles that can be flexibly positioned and dynamically altered in an open cell-chamber environment. A detailed fabrication protocol for the production of these chips using multilayer soft lithography is reported. A comprehensive computational model is also presented based on COMSOL Multiphysics software that includes both diffusion and advection of the fluid as it exits the microchannels. The results of the simulation are successfully applied to model single-channel experiments. The chip is then tested in multi-channel mode, and its ability to produce complex spatially varied concentration profiles is demonstrated. The achievement of steady state of the gradient profile in less than 5 min also allows for the dynamic variation of the profile. Finally, we apply the present chip architecture to investigate the migration of mouse neutrophils in an Interleukin-8 gradient. We report quantitatively on cell migration driven by Interleukin-8 gradient and provide migration speed distribution.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11384/6736
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