High-Throughput Single-Cell Analysis Device for Following Simultaneous Intracellular Signaling Events

Abstract

Investigating the behavior of single cells in a high-throughput manner calls for the design and realization of innovative, robust and versatile devices capable of conducting adaptable experiments to address the desired biological questions.For instance, the possibility of running experiments that compare the reaction of different cell types to a specific stimulus or the effect of a given substance at different concentrations on a specific cell type is highly desirable. One inherent challenge associated with running such experiments, however, is the existence of random uncertainty factors from experiment to experiment. Examples of this could be common measurement errors in preparing exact substance concentrations or the likeness of consecutive cell cultures.Here, we report the design and fabrication of a high-throughput microfluidic platform intended for parallel investigations on single cells upon extracellular environmental changes.We fabricated a hydrodynamic-based cell isolation microfluidic block in polydimethylsiloxane (PDMS) with two secluded entities. The comb-shaped trapping zone in each entity consists of 52 V-formed individual traps. Each trap has the dimensions of 10 μm × 10 μm and connects two complement microchannels via a 2-μm wide confinement opening. After the cell-loading step, isolated cells can undergo several chemical-rinsing steps and, in combination with fluorescence microscopy imaging, cellular response can be read and analyzed.This easy-to-operate, parallel microfluidic platform enables the simultaneous comparison of the read out between two portions of a particular cell passage to various chemical stresses or different concentrations of one specific stimulus.Moreover cellular responses from two different cell types to a known stress substance can be followed over time.We foresee that this device can be considered as the first building block of a completely integrated lab-on-a-chip platform for concentration-dependent drug screening applications and comparative single-cell data acquisition.