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Minimizing scattering-induced phase errors in differential interference contrast microscopy

Significance: Differential interference contrast (DIC) microscopes allow noninvasive in vivo observation of transparent microstructures in tissue without the use of fluorescent dyes or genetic modification. We show how to modify a DIC microscope to measure the sample phase distribution accurately an...

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Autores principales: Takano, Wataru, Shibata, Shuhei, Hagen, Nathan, Matsuda, Masaru, Otani, Yukitoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Society of Photo-Optical Instrumentation Engineers 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7734411/
https://www.ncbi.nlm.nih.gov/pubmed/33319525
http://dx.doi.org/10.1117/1.JBO.25.12.123703
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author Takano, Wataru
Shibata, Shuhei
Hagen, Nathan
Matsuda, Masaru
Otani, Yukitoshi
author_facet Takano, Wataru
Shibata, Shuhei
Hagen, Nathan
Matsuda, Masaru
Otani, Yukitoshi
author_sort Takano, Wataru
collection PubMed
description Significance: Differential interference contrast (DIC) microscopes allow noninvasive in vivo observation of transparent microstructures in tissue without the use of fluorescent dyes or genetic modification. We show how to modify a DIC microscope to measure the sample phase distribution accurately and in real-time even deep inside sample tissue. Aim: Our aim is to improve the DIC microscope’s phase measurement to remove the phase bias that occurs in the presence of strong scattering. Approach: A quarter-wave plate was added in front of the polarization camera, allowing a modified phase calculation to incorporate all four polarization orientation angles (0 deg, 45 deg, 90 deg, and 135 deg) captured simultaneously by the polarization camera, followed by deconvolution. Results: We confirm that the proposed method reduces phase measurement error in the presence of scattering and demonstrate the method using in vivo imaging of a beating heart inside a medaka egg and the whole-body blood circulation in a young medaka fish. Conclusions: Modifying a polarization-camera DIC microscope with a quarter-wave plate allows users to image deep inside samples without phase bias due to scattering effects.
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spelling pubmed-77344112020-12-16 Minimizing scattering-induced phase errors in differential interference contrast microscopy Takano, Wataru Shibata, Shuhei Hagen, Nathan Matsuda, Masaru Otani, Yukitoshi J Biomed Opt Special Series on Biomedical Imaging and Sensing Significance: Differential interference contrast (DIC) microscopes allow noninvasive in vivo observation of transparent microstructures in tissue without the use of fluorescent dyes or genetic modification. We show how to modify a DIC microscope to measure the sample phase distribution accurately and in real-time even deep inside sample tissue. Aim: Our aim is to improve the DIC microscope’s phase measurement to remove the phase bias that occurs in the presence of strong scattering. Approach: A quarter-wave plate was added in front of the polarization camera, allowing a modified phase calculation to incorporate all four polarization orientation angles (0 deg, 45 deg, 90 deg, and 135 deg) captured simultaneously by the polarization camera, followed by deconvolution. Results: We confirm that the proposed method reduces phase measurement error in the presence of scattering and demonstrate the method using in vivo imaging of a beating heart inside a medaka egg and the whole-body blood circulation in a young medaka fish. Conclusions: Modifying a polarization-camera DIC microscope with a quarter-wave plate allows users to image deep inside samples without phase bias due to scattering effects. Society of Photo-Optical Instrumentation Engineers 2020-12-14 2020-12 /pmc/articles/PMC7734411/ /pubmed/33319525 http://dx.doi.org/10.1117/1.JBO.25.12.123703 Text en © 2020 The Authors https://creativecommons.org/licenses/by/4.0/ Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
spellingShingle Special Series on Biomedical Imaging and Sensing
Takano, Wataru
Shibata, Shuhei
Hagen, Nathan
Matsuda, Masaru
Otani, Yukitoshi
Minimizing scattering-induced phase errors in differential interference contrast microscopy
title Minimizing scattering-induced phase errors in differential interference contrast microscopy
title_full Minimizing scattering-induced phase errors in differential interference contrast microscopy
title_fullStr Minimizing scattering-induced phase errors in differential interference contrast microscopy
title_full_unstemmed Minimizing scattering-induced phase errors in differential interference contrast microscopy
title_short Minimizing scattering-induced phase errors in differential interference contrast microscopy
title_sort minimizing scattering-induced phase errors in differential interference contrast microscopy
topic Special Series on Biomedical Imaging and Sensing
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7734411/
https://www.ncbi.nlm.nih.gov/pubmed/33319525
http://dx.doi.org/10.1117/1.JBO.25.12.123703
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