Continuous spectroscopic measurements of photo-stimulated release of molecules by nanomachines in a single living cell

Abstract

The first continuous, real-time spectroscopic monitoring of a photo-driven cargo delivery event from a mesoporous silica-based nanocarrier inside a single living cell is reported. By chemically attaching azobenzene molecules inside the 3 nm pore channels of mesoporous silica nanoparticles (∼70 nm diameter), the escape of the cargo molecule [propidium iodide (PI)] from the pore is prevented in the dark but is facilitated by the light-driven isomerization motion. Real-time spectroscopic measurements of a single cell uncover intermediate processes that occur during this intracellular delivery event, from nanomachine activation to the release of PI into the cytosol and to PI's eventual intercalation with nuclear DNA. Changes in PI's fluorescence intensity and the hypsochromic shift of the band maxima are used to identify the local environment of the fluorophore that is being observed in the cell. The ability to precisely initiate a chemical event inside an individual cell and continuously monitor the subsequent biological responses will enhance our understanding of intracellular process upon drug, protein and nucleic acid delivery.