Por favor, use este identificador para citar o enlazar a este item:
http://hdl.handle.net/10261/339065
COMPARTIR / EXPORTAR:
SHARE CORE BASE | |
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
Título: | Determination and localization of specific proteins in individual ARPE-19 cells by single cell and laser ablation ICP-MS using iridium nanoclusters as label |
Autor: | Menero-Valdés, Paula; Lores-Padín, Ana; Fernández-Vega, Beatriz; Quarles Jr., C. Derrick; García, Montserrat; González-Iglesias, Héctor CSIC ORCID ; Pereiro, Rosario | Palabras clave: | Retinal pigment epithelial cells Iridium nanoclusters Metal-labelled immunoprobe sc-ICP-MS LA-ICP-MS Biomaging |
Fecha de publicación: | feb-2023 | Editor: | Elsevier BV | Citación: | Talanta 253: 123974 (2023) | Resumen: | Single cell-inductively coupled plasma-mass spectrometry (sc-ICP-MS) and laser ablation (LA)-ICP-MS have been complementary employed to develop a comprehensive study of APOE and claudin-1 expression in ARPE-19 cells submitted to a glucose treatment (100 mM, 48 h) that induces oxidative stress conditions. Results were compared with control cells. The determination of the two proteins by ICP-MS was sequentially carried out using specific immunoprobes labelled with IrNCs that offer a huge amplification (1760 ± 90 atoms of Ir on average). A novel sample introduction system, the microFAST Single Cell set-up, was employed for sc-ICP-MS analysis. This introduction system resulted in a cellular transport efficiency of 85 ± 9% for ARPE-19 cells (91 ± 5% using a PtNPs standard). After the proper immunocytochemistry protocol with the specific IrNCs immunoprobes in cell suspensions (sc-ICP-MS), the mass of APOE and claudin-1 in individual ARPE-19 cells was obtained. Average detection limits per cell by sc-ICP-MS were 0.02 fg of APOE and 3 ag of claudin-1. The results of sample analyses obtained by sc-ICP-MS were validated with commercial ELISA kits. The distribution of both target proteins in individual cells (fixated in the chamber wall) was unveiled by LA-ICP-MS. The high amplification provided by the IrNCs immunoprobes allowed the identification of APOE and claudin-1 within individual ARPE-19 cells. High resolution images were obtained using a laser spot of 2 × 2 μm. | Versión del editor: | http://dx.doi.org/10.1016/j.talanta.2022.123974 | URI: | http://hdl.handle.net/10261/339065 | DOI: | 10.1016/j.talanta.2022.123974 | Identificadores: | doi: 10.1016/j.talanta.2022.123974 issn: 0039-9140 e-issn: 1873-3573 |
Aparece en las colecciones: | (IPLA) Artículos |
Ficheros en este ítem:
Fichero | Descripción | Tamaño | Formato | |
---|---|---|---|---|
1-s2.0-S0039914022007706-main.pdf | 1,52 MB | Adobe PDF | Visualizar/Abrir |
CORE Recommender
SCOPUSTM
Citations
4
checked on 07-may-2024
Page view(s)
26
checked on 15-may-2024
Download(s)
19
checked on 15-may-2024
Google ScholarTM
Check
Altmetric
Altmetric
Este item está licenciado bajo una Licencia Creative Commons