Fabrication of single cell microarrays on a double-layered hydrogel for mitochondrial activity monitoring

Abstract

Single-cell microarrays may play an important role in understanding the heterogeneity of cell populations. A hydrogel-coated micropatterned surface was prepared to capture single cells from a mixture of cells and detect their mitochondrial activity using real-time fluorescence analysis. The double-layered hydrogel micropatterns were functionalized with RGD-containing peptides using a photomask. We were able to control the number of cells immobilized on each spot of the micropattern by controlling the size of the spots. Single cells were properly arranged on the peptide-functionalized spot with aligned squares (20 μm × 20 μm), with 68% frequency of single cells. In the single-cell micropattern, a mitochondria-targetable switch-on-type of probe was employed to discriminate between the types of cells and exhibit heterogeneous cell behavior in real-time. This single cell detection technology allows for the analysis of cell function and gene expression in various environments. © 2022 Elsevier B.V.