Chen I-Jane, Wu Tao, Hu Shuhuan
Complete Genomics Inc., San Jose, CA, USA.
Beijing Genomic Institute, Shenzhen, Guandong, China.
MethodsX. 2018 Aug 20;5:984-990. doi: 10.1016/j.mex.2018.08.008. eCollection 2018.
Here we develop a microfluidic device to generate monodispersion sub-nanoliter size droplets. Our system reaches steady state within 3 s after the flow starts and generates 100,000 droplets in 28 s with high size consistency (CV < 8%). This low cost device is composed with a microfluidic chip, 2 tubings, a collection vial, a syringe and a station; and is in the size of an iPad Mini (4" × 6" × 3/4"). In this system, all incoming reagents share the same pressure drop across the fluidic passage to generator droplets. A single source negative pressure is applied to the fluids to create the flow by a vacuum at the exit end of the device. The vacuum is generated on-site by pulling the plunger of a syringe. The position of the plunger before and after pulling determines the degree of vacuum. A fixture is used to hold the plunger after it is pulled to maintain its vacuum. Although this system loses vacuum gradually as the liquid filling in, it maintains a flow rates with the changes less than 10% and droplet sizes changes less than 2% during the course of generating 150,000 droplets. The pressure drop across the chip, the flow rates of all reagents, the droplet size and generation frequency are predictable, programmable, and reproducible. This device is designed for generating droplets for single cell genome profiling application but can be also used for digital PCR or other droplet-based applications.
在此,我们开发了一种微流控装置,用于生成单分散的亚纳升尺寸液滴。我们的系统在流动开始后3秒内达到稳态,并在28秒内生成100,000个液滴,尺寸一致性高(CV < 8%)。这种低成本装置由一个微流控芯片、2根管子、一个收集瓶、一个注射器和一个工作站组成;尺寸与iPad Mini(4英寸×6英寸×3/4英寸)相同。在该系统中,所有进入的试剂在流体通道上共享相同的压降以生成液滴。对流体施加单一源负压,通过装置出口端的真空来产生流动。通过拉动注射器的活塞在现场产生真空。拉动前后活塞的位置决定了真空度。拉动活塞后使用一个固定装置来保持其真空度。尽管随着液体填充该系统会逐渐失去真空,但在生成150,000个液滴的过程中,其流速变化小于10%,液滴尺寸变化小于2%。芯片上的压降、所有试剂的流速、液滴尺寸和生成频率都是可预测、可编程且可重复的。该装置设计用于为单细胞基因组分析应用生成液滴,但也可用于数字PCR或其他基于液滴的应用。