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Spectroscopic Evaluation of Red Blood Cells of Thalassemia Patients with Confocal Microscopy: A Pilot Study

Hemoglobinopathies represent the most common single-gene defects in the world and pose a major public health problem, particularly in tropical countries, where they occur with high frequency. Diagnosing hemoglobinopathies can sometimes be difficult due to the coexistence of different causes of anemi...

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Autores principales: Rey-Barroso, Laura, Roldán, Mónica, Burgos-Fernández, Francisco J., Gassiot, Susanna, Ruiz Llobet, Anna, Isola, Ignacio, Vilaseca, Meritxell
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7412432/
https://www.ncbi.nlm.nih.gov/pubmed/32708084
http://dx.doi.org/10.3390/s20144039
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author Rey-Barroso, Laura
Roldán, Mónica
Burgos-Fernández, Francisco J.
Gassiot, Susanna
Ruiz Llobet, Anna
Isola, Ignacio
Vilaseca, Meritxell
author_facet Rey-Barroso, Laura
Roldán, Mónica
Burgos-Fernández, Francisco J.
Gassiot, Susanna
Ruiz Llobet, Anna
Isola, Ignacio
Vilaseca, Meritxell
author_sort Rey-Barroso, Laura
collection PubMed
description Hemoglobinopathies represent the most common single-gene defects in the world and pose a major public health problem, particularly in tropical countries, where they occur with high frequency. Diagnosing hemoglobinopathies can sometimes be difficult due to the coexistence of different causes of anemia, such as thalassemia and iron deficiency, and blood transfusions, among other factors, and requires expensive and complex molecular tests. This work explores the possibility of using spectral confocal microscopy as a diagnostic tool for thalassemia in pediatric patients, a disease caused by mutations in the globin genes that result in changes of the globin chains that form hemoglobin—in pediatric patients. Red blood cells (RBCs) from patients with different syndromes of alpha-thalassemia and iron deficiency (including anemia) as well as healthy (control) subjects were analyzed under a Leica TCS SP8 confocal microscope following different image acquisition protocols. We found that diseased RBCs exhibited autofluorescence when excited at 405 nm and their emission was collected in the spectral range from 425 nm to 790 nm. Three experimental descriptors calculated from the mean emission intensities at 502 nm, 579 nm, 628 nm, and 649 nm allowed us to discriminate between diseased and healthy cells. According to the results obtained, spectral confocal microscopy could serve as a tool in the diagnosis of thalassemia.
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spelling pubmed-74124322020-08-26 Spectroscopic Evaluation of Red Blood Cells of Thalassemia Patients with Confocal Microscopy: A Pilot Study Rey-Barroso, Laura Roldán, Mónica Burgos-Fernández, Francisco J. Gassiot, Susanna Ruiz Llobet, Anna Isola, Ignacio Vilaseca, Meritxell Sensors (Basel) Letter Hemoglobinopathies represent the most common single-gene defects in the world and pose a major public health problem, particularly in tropical countries, where they occur with high frequency. Diagnosing hemoglobinopathies can sometimes be difficult due to the coexistence of different causes of anemia, such as thalassemia and iron deficiency, and blood transfusions, among other factors, and requires expensive and complex molecular tests. This work explores the possibility of using spectral confocal microscopy as a diagnostic tool for thalassemia in pediatric patients, a disease caused by mutations in the globin genes that result in changes of the globin chains that form hemoglobin—in pediatric patients. Red blood cells (RBCs) from patients with different syndromes of alpha-thalassemia and iron deficiency (including anemia) as well as healthy (control) subjects were analyzed under a Leica TCS SP8 confocal microscope following different image acquisition protocols. We found that diseased RBCs exhibited autofluorescence when excited at 405 nm and their emission was collected in the spectral range from 425 nm to 790 nm. Three experimental descriptors calculated from the mean emission intensities at 502 nm, 579 nm, 628 nm, and 649 nm allowed us to discriminate between diseased and healthy cells. According to the results obtained, spectral confocal microscopy could serve as a tool in the diagnosis of thalassemia. MDPI 2020-07-21 /pmc/articles/PMC7412432/ /pubmed/32708084 http://dx.doi.org/10.3390/s20144039 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Letter
Rey-Barroso, Laura
Roldán, Mónica
Burgos-Fernández, Francisco J.
Gassiot, Susanna
Ruiz Llobet, Anna
Isola, Ignacio
Vilaseca, Meritxell
Spectroscopic Evaluation of Red Blood Cells of Thalassemia Patients with Confocal Microscopy: A Pilot Study
title Spectroscopic Evaluation of Red Blood Cells of Thalassemia Patients with Confocal Microscopy: A Pilot Study
title_full Spectroscopic Evaluation of Red Blood Cells of Thalassemia Patients with Confocal Microscopy: A Pilot Study
title_fullStr Spectroscopic Evaluation of Red Blood Cells of Thalassemia Patients with Confocal Microscopy: A Pilot Study
title_full_unstemmed Spectroscopic Evaluation of Red Blood Cells of Thalassemia Patients with Confocal Microscopy: A Pilot Study
title_short Spectroscopic Evaluation of Red Blood Cells of Thalassemia Patients with Confocal Microscopy: A Pilot Study
title_sort spectroscopic evaluation of red blood cells of thalassemia patients with confocal microscopy: a pilot study
topic Letter
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7412432/
https://www.ncbi.nlm.nih.gov/pubmed/32708084
http://dx.doi.org/10.3390/s20144039
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