Enhanced Visualization of Erythrocytes Through Photoluminescence Using NaYbF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup> Nanoparticles

Rare-earth nanoparticles (RE-NPs), particularly NaYF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup>, have emerged as a promising class of photoluminescent probes for bioimaging and sensing applications. These nanomaterials are characterized by their ability to absorb...

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Main Authors: Vivian Torres-Vera, Lorena M. Coronado, Ana Patricia Valencia, Alejandro Von Chong, Esteban Rua, Michelle Ng, Jorge Rubio-Retama, Carmenza Spadafora, Ricardo Correa
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Biosensors
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Online Access:https://www.mdpi.com/2079-6374/15/7/396
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author Vivian Torres-Vera
Lorena M. Coronado
Ana Patricia Valencia
Alejandro Von Chong
Esteban Rua
Michelle Ng
Jorge Rubio-Retama
Carmenza Spadafora
Ricardo Correa
author_facet Vivian Torres-Vera
Lorena M. Coronado
Ana Patricia Valencia
Alejandro Von Chong
Esteban Rua
Michelle Ng
Jorge Rubio-Retama
Carmenza Spadafora
Ricardo Correa
author_sort Vivian Torres-Vera
collection DOAJ
description Rare-earth nanoparticles (RE-NPs), particularly NaYF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup>, have emerged as a promising class of photoluminescent probes for bioimaging and sensing applications. These nanomaterials are characterized by their ability to absorb low-energy photons and emit higher-energy photons through an upconversion luminescence process. This process can be triggered by continuous-wave (CW) light excitation, providing a unique optical feature that is not exhibited by native biomolecules. However, the application of upconversion nanoparticles (UCNPs) in bioimaging requires systematic optimization to maximize the signal and ensure biological compatibility. In this work, we synthesized hexagonal-phase UCNPs (average diameter: 29 ± 3 nm) coated with polyacrylic acid (PAA) and established the optimal conditions for imaging human erythrocytes. The best results were obtained after a 4-h incubation in 100 mM HEPES buffer, using a nanoparticle concentration of 0.01 mg/mL and a laser current intensity of 250–300 mA. Under these conditions, the UCNPs exhibited minimal cytotoxicity and were found to predominantly localize at the erythrocyte membrane periphery, indicating surface adsorption rather than internalization. Additionally, a machine learning model (Random Forest) was implemented that classified the photoluminescent signal with 80% accuracy and 83% precision, with the signal intensity identified as the most relevant feature. This study establishes a quantitative and validated protocol that balances signal strength with cell integrity, enabling robust and automated image analysis.
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spelling doaj-art-785ff8eb1f40427bbf92e96aefd89d3d2025-08-20T03:58:31ZengMDPI AGBiosensors2079-63742025-06-0115739610.3390/bios15070396Enhanced Visualization of Erythrocytes Through Photoluminescence Using NaYbF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup> NanoparticlesVivian Torres-Vera0Lorena M. Coronado1Ana Patricia Valencia2Alejandro Von Chong3Esteban Rua4Michelle Ng5Jorge Rubio-Retama6Carmenza Spadafora7Ricardo Correa8Biomedical Physics and Engineering Unit, Center of Cellular and Molecular Biology of Diseases (CBCMe), Instituto de Investigaciones Científicas y Servicios de Alta Tecnología (INDICASAT AIP), Panama City 1843-01103, PanamaBiomedical Physics and Engineering Unit, Center of Cellular and Molecular Biology of Diseases (CBCMe), Instituto de Investigaciones Científicas y Servicios de Alta Tecnología (INDICASAT AIP), Panama City 1843-01103, PanamaFacultad de Ingeniería, Universidad Latina de Panamá, Panama City 0823-00933, PanamaSistema Nacional de Investigación (SNI), Secretaría Nacional de Ciencia, Tecnología e Innovación, Panama City 0816-02852, PanamaSchool of Electrical Engineering, Universidad Tecnológica de Panamá, Víctor Levi Sasso Campus, Panama City 0819-07289, PanamaBiomedical Physics and Engineering Unit, Center of Cellular and Molecular Biology of Diseases (CBCMe), Instituto de Investigaciones Científicas y Servicios de Alta Tecnología (INDICASAT AIP), Panama City 1843-01103, PanamaDepartment of Chemistry in Pharmaceutical Sciences, Faculty of Pharmacy, Complutense University of Madrid, Plaza Ramon y Cajal 2, 28040 Madrid, SpainBiomedical Physics and Engineering Unit, Center of Cellular and Molecular Biology of Diseases (CBCMe), Instituto de Investigaciones Científicas y Servicios de Alta Tecnología (INDICASAT AIP), Panama City 1843-01103, PanamaBiomedical Physics and Engineering Unit, Center of Cellular and Molecular Biology of Diseases (CBCMe), Instituto de Investigaciones Científicas y Servicios de Alta Tecnología (INDICASAT AIP), Panama City 1843-01103, PanamaRare-earth nanoparticles (RE-NPs), particularly NaYF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup>, have emerged as a promising class of photoluminescent probes for bioimaging and sensing applications. These nanomaterials are characterized by their ability to absorb low-energy photons and emit higher-energy photons through an upconversion luminescence process. This process can be triggered by continuous-wave (CW) light excitation, providing a unique optical feature that is not exhibited by native biomolecules. However, the application of upconversion nanoparticles (UCNPs) in bioimaging requires systematic optimization to maximize the signal and ensure biological compatibility. In this work, we synthesized hexagonal-phase UCNPs (average diameter: 29 ± 3 nm) coated with polyacrylic acid (PAA) and established the optimal conditions for imaging human erythrocytes. The best results were obtained after a 4-h incubation in 100 mM HEPES buffer, using a nanoparticle concentration of 0.01 mg/mL and a laser current intensity of 250–300 mA. Under these conditions, the UCNPs exhibited minimal cytotoxicity and were found to predominantly localize at the erythrocyte membrane periphery, indicating surface adsorption rather than internalization. Additionally, a machine learning model (Random Forest) was implemented that classified the photoluminescent signal with 80% accuracy and 83% precision, with the signal intensity identified as the most relevant feature. This study establishes a quantitative and validated protocol that balances signal strength with cell integrity, enabling robust and automated image analysis.https://www.mdpi.com/2079-6374/15/7/396rare-earth nanoparticlesred blood cellsphotoluminescencebiocompatibility
spellingShingle Vivian Torres-Vera
Lorena M. Coronado
Ana Patricia Valencia
Alejandro Von Chong
Esteban Rua
Michelle Ng
Jorge Rubio-Retama
Carmenza Spadafora
Ricardo Correa
Enhanced Visualization of Erythrocytes Through Photoluminescence Using NaYbF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup> Nanoparticles
Biosensors
rare-earth nanoparticles
red blood cells
photoluminescence
biocompatibility
title Enhanced Visualization of Erythrocytes Through Photoluminescence Using NaYbF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup> Nanoparticles
title_full Enhanced Visualization of Erythrocytes Through Photoluminescence Using NaYbF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup> Nanoparticles
title_fullStr Enhanced Visualization of Erythrocytes Through Photoluminescence Using NaYbF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup> Nanoparticles
title_full_unstemmed Enhanced Visualization of Erythrocytes Through Photoluminescence Using NaYbF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup> Nanoparticles
title_short Enhanced Visualization of Erythrocytes Through Photoluminescence Using NaYbF<sub>4</sub>:Yb<sup>3+</sup>,Er<sup>3+</sup> Nanoparticles
title_sort enhanced visualization of erythrocytes through photoluminescence using naybf sub 4 sub yb sup 3 sup er sup 3 sup nanoparticles
topic rare-earth nanoparticles
red blood cells
photoluminescence
biocompatibility
url https://www.mdpi.com/2079-6374/15/7/396
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