A silicon single-chip microfluidics system that integrates microscale fluidic channels, an analyte segmentation device, and a nozzle for electrohydrodynamic-assisted printing is designed for hyphenation with MALDI mass spectrometry (MS) imaging. A miniaturized T-junction segments analytes into monodisperse picoliter oil-isolated compartments. The printing nozzle deposits generated droplets one-by-one into an array on a conductive substrate without splitting or coalescing. Virtually single-shot MS analysis is enabled due to the ultrasmall droplet volumes and highly localized printing. The signal-to-noise ratio indicates that detection limits at the attomole level are achieved for γ-aminobutyric acid.