Work

Determination of intracellular localization of fluorescent nanoparticles v1

Year: 2026

Type: preprint

Abstract: This protocol investigates cellular uptake mechanisms and intracellular localization of polystyrene nanoparticles (PS-NPs) in HEK 293 cells. Confocal microscopy is used to monitor time-dependent internalization, clathrin-mediated endocytosis inhibition, and vesicular trafficking. It quantitatively analyzes nanoparticle internalization and spatial distribution relative to the nucleus using image segmentation and computational analysis, and evaluates how serum protein adsorption (protein corona formation) modifies PS-NP surface charge, size, and cellular uptake. Results: Both positive and negative PS-NPs entered cells primarily by clathrin-mediated endocytosis and were rapidly trapped by lysosomes. However, amine modified positive NPs were more uptaken than negative ones.

DOI: 10.17504/protocols.io.r3l25mpbpl1y/v1

Authors: Giulia Yuri Moscatiello, Paolo Bigini, Luisa Diomede

Institution: Mario Negri Institute for Pharmacological Research

Language: English

Topic: Nanoparticle-Based Drug Delivery

Subfield: Biomaterials

Field: Materials Science

Domain: Physical Sciences

SDG: Good health and well-being

Open Access status: green

Funders: European Commission, HORIZON EUROPE Framework Programme, European Health and Digital Executive Agency

Awards: Platform Optimisation To Enable NanomaTerIAL safety assessment for rapid commercialisation, G7598473398