Targeted fluorescent lipid microparticles for quantitative measurement of phagosomal pH

Phagosomal acidification plays a pivotal role in pathogen destruction and immune signaling, yet tools capable of reporting these biochemical changes with spatial and mechanistic precision remain scarce. A modular biosensing strategy is presented in which BODIPY-derived hydrophilic fluorophores are conjugated to phospholipids and incorporated at the surface of targeted lipid microparticles. These soft and biomimetic particles combine receptor-specific uptake with ratiometric fluorescence readouts, enabled by pairing pH-responsive dyes at the interface with a pH-invariant reference probe in the core. Following Fcγ receptor-mediated internalization by macrophages, the particles deliver real-time insight into the onset and progression of phagosomal acidification. This versatile platform provides a direct means to couple defined surface recognition events to intracellular pH measurements, offering new opportunities to unravel how particle identity and ligand presentation modulate phagosomal physiology.