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Label-free bioanalysis of Leishmania infantum using refractive index tomography with partially coherent illumination

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2019

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WILEY‐VCH Verlag GmbH
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In this work, we report the use of refractive index (RI) tomography for quantitative analysis of unstained DH82 cell line infected with Leishmania infantum. The cell RI is reconstructed by using a modality of optical diffraction tomography technique that employs partially coherent illumination, thus enabling inherent compatibility with conventional wide-field microscopes. The experimental results demonstrate that the cell dry mass concentration (DMC) obtained from the RI allows for reliable detection and quantitative characterization of the infection and its temporal evolution. The RI provides important insight for studying morphological changes, particularly membrane blebbing linked to an apoptosis (cell death) process induced by the disease. Moreover, the results evidence that infected DH82 cells exhibit a higher DMC than healthy samples. These findings open up promising perspectives for clinical diagnosis of Leishmania.

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© 2019 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim. The Ministerio de Economía y Competitividad is acknowledged for funding the project TEC2014-57394-P. This research was partially supported through a Santander-UCM 2018 project (PR75/18-21567) and by a grant from Comunidad de Madrid (Spain) PLATESA (S2013/ABI-2906). Alicia Mas was supported by a fellowship from Complutense University of Madrid-Santander.

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