Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer.

Détails

ID Serval
serval:BIB_0E05E28FB41D
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer.
Périodique
Cytometry. Part A
Auteur(s)
Rappaz B., Barbul A., Emery Y., Korenstein R., Depeursinge C., Magistretti P.J., Marquet P.
ISSN
1552-4930 (Electronic)
ISSN-L
1552-4922
Statut éditorial
Publié
Date de publication
10/2008
Peer-reviewed
Oui
Volume
73
Numéro
10
Pages
895-903
Langue
anglais
Notes
Publication types: Comparative Study ; Evaluation Studies ; Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Résumé
Red blood cell (RBC) parameters such as morphology, volume, refractive index, and hemoglobin content are of great importance for diagnostic purposes. Existing approaches require complicated calibration procedures and robust cell perturbation. As a result, reference values for normal RBC differ depending on the method used. We present a way for measuring parameters of intact individual RBCs by using digital holographic microscopy (DHM), a new interferometric and label-free technique with nanometric axial sensitivity. The results are compared with values achieved by conventional techniques for RBC of the same donor and previously published figures. A DHM equipped with a laser diode (lambda = 663 nm) was used to record holograms in an off-axis geometry. Measurements of both RBC refractive indices and volumes were achieved via monitoring the quantitative phase map of RBC by means of a sequential perfusion of two isotonic solutions with different refractive indices obtained by the use of Nycodenz (decoupling procedure). Volume of RBCs labeled by membrane dye Dil was analyzed by confocal microscopy. The mean cell volume (MCV), red blood cell distribution width (RDW), and mean cell hemoglobin concentration (MCHC) were also measured with an impedance volume analyzer. DHM yielded RBC refractive index n = 1.418 +/- 0.012, volume 83 +/- 14 fl, MCH = 29.9 pg, and MCHC 362 +/- 40 g/l. Erythrocyte MCV, MCH, and MCHC achieved by an impedance volume analyzer were 82 fl, 28.6 pg, and 349 g/l, respectively. Confocal microscopy yielded 91 +/- 17 fl for RBC volume. In conclusion, DHM in combination with a decoupling procedure allows measuring noninvasively volume, refractive index, and hemoglobin content of single-living RBCs with a high accuracy.

Mots-clé
Electric Impedance, Erythrocyte Indices, Erythrocyte Volume, Erythrocytes/cytology, Holography, Humans, Microscopy, Confocal/methods, Reference Values, Sensitivity and Specificity, Signal Processing, Computer-Assisted
Pubmed
Web of science
Open Access
Oui
Création de la notice
13/02/2009 14:51
Dernière modification de la notice
08/05/2019 14:25
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