Despite the precise controllability of droplet samples in digital microfluidic(DMF)systems,their capability in isolating single cells for long-time culture is still limited:typically,only a few cells can be captured o...Despite the precise controllability of droplet samples in digital microfluidic(DMF)systems,their capability in isolating single cells for long-time culture is still limited:typically,only a few cells can be captured on an electrode.Although fabricating small-sized hydrophilic micropatches on an electrode aids single-cell capture,the actuation voltage for droplet transportation has to be significantly raised,resulting in a shorter lifetime for the DMF chip and a larger risk of damaging the cells.In this work,a DMF system with 3D microstructures engineered on-chip is proposed to form semi-closed micro-wells for efficient single-cell isolation and long-time culture.Our optimum results showed that approximately 20%of the micro-wells over a 30×30 array were occupied by isolated single cells.In addition,low-evaporation-temperature oil and surfactant aided the system in achieving a low droplet actuation voltage of 36V,which was 4 times lower than the typical 150 V,minimizing the potential damage to the cells in the droplets and to the DMF chip.To exemplify the technological advances,drug sensitivity tests were run in our DMF system to investigate the cell response of breast cancer cells(MDA-MB-231)and breast normal cells(MCF-10A)to a widely used chemotherapeutic drug,Cisplatin(Cis).The results on-chip were consistent with those screened in conventional 96-well plates.This novel,simple and robust single-cell trapping method has great potential in biological research at the single cell level.展开更多
基金This work was supported by the FDCT110/2016/A3AMSV SKL Fund from the Macao Science and Technology Development Fund(FDCT)by MYRG2017-00022-AMSV and MYRG2018-00114-AMSV from the University of Macao.
文摘Despite the precise controllability of droplet samples in digital microfluidic(DMF)systems,their capability in isolating single cells for long-time culture is still limited:typically,only a few cells can be captured on an electrode.Although fabricating small-sized hydrophilic micropatches on an electrode aids single-cell capture,the actuation voltage for droplet transportation has to be significantly raised,resulting in a shorter lifetime for the DMF chip and a larger risk of damaging the cells.In this work,a DMF system with 3D microstructures engineered on-chip is proposed to form semi-closed micro-wells for efficient single-cell isolation and long-time culture.Our optimum results showed that approximately 20%of the micro-wells over a 30×30 array were occupied by isolated single cells.In addition,low-evaporation-temperature oil and surfactant aided the system in achieving a low droplet actuation voltage of 36V,which was 4 times lower than the typical 150 V,minimizing the potential damage to the cells in the droplets and to the DMF chip.To exemplify the technological advances,drug sensitivity tests were run in our DMF system to investigate the cell response of breast cancer cells(MDA-MB-231)and breast normal cells(MCF-10A)to a widely used chemotherapeutic drug,Cisplatin(Cis).The results on-chip were consistent with those screened in conventional 96-well plates.This novel,simple and robust single-cell trapping method has great potential in biological research at the single cell level.
