Monitoring Nanoparticle Interaction with Murine Breast Cancer Cells Using Multimodal Fluorescence Lifetime Microscopy

To investigate drug delivery in cancer therapy, we integrate fluorescence lifetime measurements, microspectrometry, and confocal laser scanning microscopy to track the uptake of inorganic–organic hybrid nanoparticles (IOH-NPs) by breast cancer cells over incubation periods ranging from 2 to 24 h. Non-radiative energy transfer (FRET) from the LysoTracker Green to the IOH-NPs confirms their lysosomal localization and possibly improves their optical excitation. Beyond the resolution limits of light and electron microscopy, fluorescence lifetime kinetics—including FRET—can thus reveal the nanoscale cellular localization of IOH-NPs and guide the optimization of fluorescence excitation. Here, we extend optical microscopy into a fifth dimension—picosecond fluorescence decay times—complementing 3D spatial and spectral information, establishing lifetime measurements as a versatile tool to study nanoparticle uptake in cancer therapy.