Super‐Resolution Microscopy Unveils Dynamic Heterogeneities in Nanoparticle Protein Corona

Abstract

The adsorption of serum proteins, leading to the formation of a biomolecular corona, is a key determinant of the biological identity of nanoparticles in vivo. Therefore, gaining knowledge on the formation, composition, and temporal evolution of the corona is of utmost importance for the development of nanoparticle-based therapies. Here, it is shown that the use of super-resolution optical microscopy enables the imaging of the protein corona on mesoporous silica nanoparticles with single protein sensitivity. Particle-by-particle quantification reveals a significant heterogeneity in protein absorption under native conditions. Moreover, the diversity of the corona evolves over time depending on the surface chemistry and degradability of the particles. This paper investigates the consequences of protein adsorption for specific cell targeting by antibody-functionalized nanoparticles providing a detailed understanding of corona-activity relations. The methodology is widely applicable to a variety of nanostructures and complements the existing ensemble approaches for protein corona study.

Super-resolution microscopy as a powerful tool to study complex synthetic materials

“…This work unveiled for the first time heterogeneity in the number of proteins attached to the different nanoparticles of the same sample that could only be revealed thanks to the nanometric resolution of the technique. This information is of relevant interest when studying the functionality of the nanoparticles in vivo, as different protein corona may result in different performance of each individual nanoparticle 139 . STORM has also been used to study the penetration of different serum proteins within porous silica nanoparticles 140 .…”

Tracking the DNA complexation state of pBAE polyplexes in cells with super resolution microscopy

“…To objectively identify and quantify polyplexes, the list of localizations of the dSTORM images were exported and analysed in a Matlab script previously described by Feiner-Gracia et al 45 Briefly, the pDNA localizations were clustered using a mean shift algorithm with a bandwidth of 80 nm. An ellipse was fitted on the obtained clusters in order to filter out clusters with an aspect ratio higher than 5.…”

Two‐Photon‐Excited Silica and Organosilica Nanoparticles for Spatiotemporal Cancer Treatment

“…We will consider four types of organosilica nanoplatforms for TPE biomedical applications: mesoporous silica nanoparticles (MSNs), mesoporous organosilica nanoparticles (MONs), periodic mesoporous organosilica (PMO) NPs, and bridge silsesquioxanes NPs (BS NPs) ( Figure a–d). First, MSNs (Figure a) are typically obtained by the sol–gel process of silica precursors such as tetraethoxysilane (TEOS) under basic catalysis in the presence of an aqueous cetyltrimethylammonium bromide template.…”