Raman-Activated Droplet Sorting (RADS) for Label-Free High-Throughput Screening of Microalgal Single-Cells.

Raman-activated cell sorting (RACS) has attracted increasing interest, yet throughput remains one major factor limiting its broader application. Here we present an integrated Raman-activated droplet sorting (RADS) microfluidic system for functional screening of live cells in a label-free and high-throughput manner, by employing AXT-synthetic industrial microalga Haematococcus pluvialis (H. pluvialis) as a model. Raman microspectroscopy analysis of individual cells is carried out prior to their microdroplet encapsulation, which is then directly coupled to DEP-based droplet sorting. To validate the system, H. pluvialis cells containing different levels of AXT were mixed and underwent RADS. Those AXT-hyperproducing cells were sorted with an accuracy of 98.3%, an enrichment ratio of eight folds, and a throughput of ∼260 cells/min. Of the RADS-sorted cells, 92.7% remained alive and able to proliferate, which is equivalent to the unsorted cells. Thus, the RADS achieves a much higher throughput than existing RACS systems, preserves the vitality of cells, and facilitates seamless coupling with downstream manipulations such as single-cell sequencing and cultivation.

On-demand control of microfluidic flow via capillary-tuned solenoid microvalve suction.

A simple, low-cost and on-demand microfluidic flow controlling platform was developed based on a unique capillary-tuned solenoid microvalve suction effect without any outer pressure source. The suction effect was innovatively employed as a stable and controllable driving force for the manipulation of the microfluidic system by connecting a piece of capillary between the microvalve and the microfluidic chip, which caused significant hydrodynamic resistance differences among the solenoid valve ports and changed the flowing mode inside the valve. The volume of sucked liquid could be controlled from microliters even down to picoliters either by decreasing the valve energized duration (from a maximum energized duration to the valve response time of 20 ms) or by increasing the inserted capillary length (i.e., its hydrodynamic resistance). Several important microfluidic unit operations such as cell/droplet sorting and on-demand size-controllable droplet generation have been demonstrated on the developed platform and both simulations and experiments confirmed that this platform has good controllability and stability.