Harnessing HetHydrogel: A Universal Platform to Dropletize Single‐Cell Multiomics

Abstract A universal platform is developed for dropletizing single cell plate‐based multiomic assays, consisting of three main pillars: a miniaturized open Heterogeneous Hydrogel reactor (abbreviated HetHydrogel) for multi‐step biochemistry, its tunable permeability that allows Tn5 tagmentation, and single cell droplet barcoding. Through optimizing the HetHydrogel manufacturing procedure, the chemical composition, and cell permeation conditions, simultaneous high‐throughput mitochondrial DNA genotyping and chromatin profiling at the single‐cell level are demonstrated using a mixed‐species experiment. This platform offers a powerful way to investigate the genotype‐phenotype relationships of various mtDNA mutations in biological processes. The HetHydrogel platform is believed to have the potential to democratize droplet technologies, upgrading a whole range of plate‐based single cell assays to high throughput format.