26 June 2017 Digital holographic microscopy as a technique to monitor macrophages infected by leishmania
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Abstract
The Digital Holographic Microscopy in Transmission technique (DHM) is considered a useful tool in the noninvasive quantifying of transparent biological objects like living cells. In this work, we propose this technique to study and to monitor control macrophages infected by Leishmania (mouse lineJ774.A1). When the promastigotes enter in contact with healthy macrophages, they got phagocytosed and latterly confined in the formed parasitophorous vacuole. These processes change the morphology and density of the host macrophage. Both parameters can be measured in a label-free analysis of cells with the aid of the DHM technique. Our technique begins with the optical record of the holograms using a modified Mach-Zehnder interferometer and the reconstruction of the complex optical field transmitted by macrophages. In the latter point, we employ the angular spectrum algorithm. With the complex optical field reconstruction, we compute the field amplitude and the phase difference maps, which leads to describe one morphological characterization for the samples. Using phase difference maps is possible to measure internal variations for the integral refractive index, estimating the infection level of macrophages. Through the changes in the integral refractive index, it is also possible to describe and quantify in two different states the evolution of the infection. With these results some parameters of cells have been quantified, making the DHM technique a viable tool for diagnosis of biological samples under the presence of some pathogen.
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E. Mendoza-Rodríguez, E. Mendoza-Rodríguez, C. Organista-Castelblanco, C. Organista-Castelblanco, M. Camacho, M. Camacho, F. Monroy-Ramírez, F. Monroy-Ramírez, } "Digital holographic microscopy as a technique to monitor macrophages infected by leishmania", Proc. SPIE 10333, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials III, 103330P (26 June 2017); doi: 10.1117/12.2271955; https://doi.org/10.1117/12.2271955
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