Facile One Pot Synthesis of Hybrid Core-Shell Silica-Based Sensors for Live Imaging of Dissolved Oxygen and Hypoxia Mapping in 3D cell models

Fuente: arXiv
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Main Authors: Iuele, Helena, Forciniti, Stefania, Onesto, Valentina, Colella, Francesco, Siciliano, Anna Chiara, Chandra, Anil, Nobile, Concetta, Gigli, Giuseppe, del Mercato, Loretta L.
Format: Preprint
Published: 2024
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author Iuele, Helena
Forciniti, Stefania
Onesto, Valentina
Colella, Francesco
Siciliano, Anna Chiara
Chandra, Anil
Nobile, Concetta
Gigli, Giuseppe
del Mercato, Loretta L.
author_facet Iuele, Helena
Forciniti, Stefania
Onesto, Valentina
Colella, Francesco
Siciliano, Anna Chiara
Chandra, Anil
Nobile, Concetta
Gigli, Giuseppe
del Mercato, Loretta L.
contents Fluorescence imaging allows for non-invasively visualizing and measuring key physiological parameters like pH and dissolved oxygen. In our work, we created two ratiometric fluorescent microsensors designed for accurately tracking dissolved oxygen levels in 3D cell cultures. We developed a simple and cost-effective method to produce hybrid core-shell silica microparticles that are biocompatible and versatile. These sensors incorporate oxygen-sensitive probes (Ru(dpp) or PtOEP) and reference dyes (RBITC or A647 NHS-Ester). SEM analysis confirmed efficient loading and distribution of the sensing dye on the outer shell. Fluorimetric and CLSM tests demonstrated the sensors' reversibility and high sensitivity to oxygen, even when integrated into 3D scaffolds. Aging and bleaching experiments validated the stability of our hybrid core-shell silica microsensors for 3D monitoring. The Ru(dpp)-RBITC microparticles showed the most promising performance, especially in a pancreatic cancer model using alginate microgels. By employing computational segmentation, we generated 3D oxygen maps during live cell imaging, revealing oxygen gradients in the extracellular matrix and indicating a significant decrease in oxygen levels characteristic of solid tumors. Notably, after 12 hours, the oxygen concentration dropped to a hypoxic level of PO2 2.7 +/- 0.1%.
format Preprint
id arxiv_https___arxiv_org_abs_2410_17797
institution arXiv
publishDate 2024
record_format arxiv
spellingShingle Facile One Pot Synthesis of Hybrid Core-Shell Silica-Based Sensors for Live Imaging of Dissolved Oxygen and Hypoxia Mapping in 3D cell models
Iuele, Helena
Forciniti, Stefania
Onesto, Valentina
Colella, Francesco
Siciliano, Anna Chiara
Chandra, Anil
Nobile, Concetta
Gigli, Giuseppe
del Mercato, Loretta L.
Medical Physics
Chemical Physics
Fluorescence imaging allows for non-invasively visualizing and measuring key physiological parameters like pH and dissolved oxygen. In our work, we created two ratiometric fluorescent microsensors designed for accurately tracking dissolved oxygen levels in 3D cell cultures. We developed a simple and cost-effective method to produce hybrid core-shell silica microparticles that are biocompatible and versatile. These sensors incorporate oxygen-sensitive probes (Ru(dpp) or PtOEP) and reference dyes (RBITC or A647 NHS-Ester). SEM analysis confirmed efficient loading and distribution of the sensing dye on the outer shell. Fluorimetric and CLSM tests demonstrated the sensors' reversibility and high sensitivity to oxygen, even when integrated into 3D scaffolds. Aging and bleaching experiments validated the stability of our hybrid core-shell silica microsensors for 3D monitoring. The Ru(dpp)-RBITC microparticles showed the most promising performance, especially in a pancreatic cancer model using alginate microgels. By employing computational segmentation, we generated 3D oxygen maps during live cell imaging, revealing oxygen gradients in the extracellular matrix and indicating a significant decrease in oxygen levels characteristic of solid tumors. Notably, after 12 hours, the oxygen concentration dropped to a hypoxic level of PO2 2.7 +/- 0.1%.
title Facile One Pot Synthesis of Hybrid Core-Shell Silica-Based Sensors for Live Imaging of Dissolved Oxygen and Hypoxia Mapping in 3D cell models
topic Medical Physics
Chemical Physics
url https://arxiv.org/abs/2410.17797