High-Resolution Imaging of Intracellular Calcium Fluctuations Caused by Oscillating Microbubbles

Abstract

Ultrasound insonification of microbubbles can locally enhance drug delivery, but the microbubble-cell interaction remains poorly understood. Because intracellular calcium (Ca_i²⁺) is a key cellular regulator, unraveling the Ca_i²⁺ fluctuations caused by an oscillating microbubble provides crucial insight into the underlying bio-effects. Therefore, we developed an optical imaging system at nanometer and nanosecond resolution that can resolve Ca_i²⁺ fluctuations and microbubble oscillations. Using this system, we clearly distinguished three Ca_i²⁺ uptake profiles upon sonoporation of endothelial cells, which strongly correlated with the microbubble oscillation amplitude, severity of sonoporation and opening of cell-cell contacts. We found a narrow operating range for viable drug delivery without lethal cell damage. Moreover, adjacent cells were affected by a calcium wave propagating at 15 μm/s. With the unique optical system, we unraveled the microbubble oscillation behavior required for drug delivery and Ca_i²⁺ fluctuations, providing new insight into the microbubble-cell interaction to aid clinical translation.

Keywords: Cell–cell contact opening; Confocal microscopy; Drug delivery; High-speed imaging; Intracellular calcium; Microbubbles; Sonoporation; Ultrasound.