Retrieval of cells from interfaces after microscopic observation for follow-up “omics” measurements is a critical but often overlooked step that limits many cell-based assays. In this work, photodegradable hydrogels are developed to non-destructively recover cells from screening interfaces in order to couple an observed phenotype with genomic information. Polyethylene glycol (PEG)-based hydrogels are formed using thiol-acrylate Michael-type addition reactions to first immobilize individual bacteria cells from a heterogenous population for culture and/or microscopic observation. PEG molecules are modified with <i>o</i>-nitrobenzyl chromophores, allowing for controlled degradation of hydrogel areas with high spatial precision using a patterned illumination tool and a 365 nm light source. Patterned light exposure triggers the efficient release of targeted bacteria cells into solution for retrieval, enrichment, and sequencing. Using this methodology, photodegradable hydrogels have been developed as a light-responsive membrane for cell culture and retrieval from a microwell platform designed to screen and discover microbial interactions in root and soil microbiomes. This approach can also be used to recover and sequence strains showing rare or unique function during high-throughput screening of transposon mutant libraries. We aim to further develop this method towards precision extraction at the single cell level, and expect that the approach will be useful in a variety of high-throughput screening applications that aim to connect cellular function with genomic, proteomic, or metabolomic measurements.

Photodegradable hydrogels for retrieval of bacteria cells from screening interfaces

2019 Fall National Meeting

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