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

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. [Display omitted] • We fabricated single-cell microarrays on a PEG hydrogel-coated surface, allowing real-time analysis of single cells. • Single cells were properly arranged on the peptide-functionalized spot in pattern, with 68% frequency. • A mitochondria-targetable was employed to discriminate between the types of cells and exhibit heterogeneous cell behavior. • The mitochondrial activity was monitored by fluorescence analysis using a single cell array. • The signals from MDA-MB-231 cells showed a stronger red intensity, whereas NIH-3T3 cells exhibited a weaker red intensity. [ABSTRACT FROM AUTHOR]