Image-Based Single-Cell Sorting via Dual-Photopolymerized Microwell Arrays

We present a simple and cost-effective single-cell sorting method using two sequential photopolymerization steps that enables sorting based upon imaged phenotypes. The first photopolymerization step uses a thiolene-based resin with minimal autofluorescence to create an array of microwells to capture cells. The second photopolymerization uses (poly)ethylene glycol diacrylate to encapsulate undesired cells in a hydrogel, allowing for retrieval of the desired cell population using simple washing. We quantitatively characterize the method using fluorescently labeled cells and then applied the method to isolate cells based upon imaged fluorescence localization. The method is readily transferrable to other laboratories and will provide a facile route to sorting of cells based on imaged phenotypes.