Regensburg 2025 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 22: Bioimaging
BP 22.2: Vortrag
Mittwoch, 19. März 2025, 15:30–15:45, H46
3D Single-Nanoparticle Tracking with Fluorescence Lifetime Imaging for Investigating Lipid Nanoparticles Endosomal Pathways — •Thomas Kellerer1,2, Judith A. Müller2, Tanja Grawert1, Lukas Moser1, Joachim O. Rädler2, and Thomas Hellerer1 — 1Multiphoton Imaging Lab, Munich University of Applied Sciences, 80335 Munich, Germany — 2Faculty of Physics and Center for NanoScience, Ludwig Maximilians-University, 80539 Munich, Germany
Lipid nanoparticles (LNPs) are vital for delivering mRNA in drug delivery systems, but the kinetics and intracellular pathways of their cargo release remain often unclear. To address this, we developed a microscopy technique to track single nanoparticles in 3D over extended periods. This approach integrates a lock-in amplifier for simultaneous image based 3D tracking and fluorescence lifetime measurement, enabling analysis of the microenvironment, such as pH changes. Achieving a frame rate of 7.6 fps at 1024x1024 resolution and lifetimes measured within 102 ns, our method provides novel insights into LNP dynamics and endosomal acidification in live cells. This technique was applied to study the acidification kinetics of endosomes during transfection. By measuring pH changes in real time, we provided insights into the intracellular behavior of LNPs and their role in mRNA delivery. This approach establishes a new standard for tracking nanoparticles and analyzing their microenvironments with high spatial and temporal resolution.
Keywords: Lipid Nanoparticles; Fluorescence Lifetime Imaging Microscopy; Single Particle Tracking; Microenvironmental Sensing; Endosomal Pathway