Open Microfluidic Probe Enabled Regional Surface Modification of Single Cells by Click Chemistry

Abstract Precise subcellular modification of single-cell surfaces is essential for studying cell polarization, localized signaling, and targeted pharmacology. However, existing methods typically produce isotropic modifications with limited spatial control, restricting the study of subcellular processes. Herein, we present a microfluidic probe (MFP)-based strategy that combines metabolic glycoengineering (MGE) with click chemistry for regional surface modification of living cells (U87 and U251). Hydrodynamic flow confinement (HFC) enables two chemically distinct regions to be delivered to opposite sides of a single cell, allowing dual-color fluorescent labeling and localized drug conjugation via strain-promoted azide−alkyne cycloaddition (SPAAC). Using monomethyl auristatin F (MMAF) as a model, we demonstrate spatially resolved cytoskeletal disruption and cell contraction. This biocompatible, spatially programmable approach provides a versatile platform for subcellular chemical biology and precision delivery.