3D Morphometry of Red Blood Cells by Digital Holography

Year: 2014

Authors: Memmolo P., Miccio L., Merola F., Gennari O., Netti P.A., Ferraro P.

Autors Affiliation: Center for Advanced Biomaterials for Health Care@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci
53, Napoli 80125, Italy;
CNR—Istituto Nazionale di Ottica, Via Campi Flegrei 34, Pozzuoli (NA) I-80078, Italy;
Department of Materials and Production Engineering University of Naples Federico II, P.le Tecchio 80, 80125 Napoli,
Italy

Abstract: Three dimensional (3D) morphometric analysis of flowing and not-adherent cells is an important aspect for diagnostic purposes. However, diagnostics tools need to be quantitative, label-free and, as much as possible, accurate. Recently, a simple holographic approach, based on shape from silhouette algorithm, has been demonstrated for accurate calculation of cells biovolume and displaying their 3D shapes. Such approach has been adopted in combination with holographic optical tweezers and successfully applied to cells with convex shape. Nevertheless, unfortunately, the method fails in case of specimen with concave surfaces. Here, we propose an effective approach to achieve correct 3D shape measurement that can be extended in case of cells having concave surfaces, thus overcoming the limit of the previous technique. We prove the new procedure for healthy red blood cells (RBCs) (i.e., discocytes) having a concave surface in their central region. Comparative analysis of experimental results with a theoretical 3D geometrical model of RBC is discussed in order to evaluate accuracy of the proposed approach. Finally, we show that the method can be also useful to classify, in terms of morphology, different varieties of RBCs. (c) 2014 International Society for Advancement of Cytometry

Journal/Review: CYTOMETRY PART A

Volume: 85A (12)      Pages from: 1030  to: 1036

KeyWords: red blood cell; morphological analysis; digital holography; microfluidic; optical tweezers
DOI: 10.1002/cyto.a.22570

Citations: 100
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