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Adaptive phase contrast microscopy to compensate for the meniscus effect
Phase contrast is one of the most important microscopic methods for making visible transparent, unstained cells. Cell cultures are often cultivated in microtiter plates, consisting of several cylindrical wells. The surface tension of the culture medium forms a liquid lens within the well, causing ph...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10082838/ https://www.ncbi.nlm.nih.gov/pubmed/37031241 http://dx.doi.org/10.1038/s41598-023-32917-6 |
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author | Nienhaus, Florian Piotrowski, Tobias Nießing, Bastian König, Niels Schmitt, Robert H. |
author_facet | Nienhaus, Florian Piotrowski, Tobias Nießing, Bastian König, Niels Schmitt, Robert H. |
author_sort | Nienhaus, Florian |
collection | PubMed |
description | Phase contrast is one of the most important microscopic methods for making visible transparent, unstained cells. Cell cultures are often cultivated in microtiter plates, consisting of several cylindrical wells. The surface tension of the culture medium forms a liquid lens within the well, causing phase contrast conditions to fail in the more curved edge areas, preventing cell observation. Adaptive phase contrast microscopy is a method to strongly increase the observable area by optically compensating for the meniscus effect. The microscope’s condenser annulus is replaced by a transmissive LCD to allow dynamic changes. A deformable, liquid-filled prism is placed in the illumination path. The prism’s surface angle is adaptively inclined to refract transmitted light so that the tangential angle of the liquid lens can be compensated. Besides the observation of the phase contrast image, a beam splitter allows to simultaneously view condenser annulus and phase ring displacement. Algorithms analyze the displacement to dynamically adjust the LCD and prism to guarantee phase contrast conditions. Experiments show a significant increase in observable area, especially for small well sizes. For 96-well-plates, more than twelve times the area can be examined under phase contrast conditions instead of standard phase contrast microscopy. |
format | Online Article Text |
id | pubmed-10082838 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-100828382023-04-10 Adaptive phase contrast microscopy to compensate for the meniscus effect Nienhaus, Florian Piotrowski, Tobias Nießing, Bastian König, Niels Schmitt, Robert H. Sci Rep Article Phase contrast is one of the most important microscopic methods for making visible transparent, unstained cells. Cell cultures are often cultivated in microtiter plates, consisting of several cylindrical wells. The surface tension of the culture medium forms a liquid lens within the well, causing phase contrast conditions to fail in the more curved edge areas, preventing cell observation. Adaptive phase contrast microscopy is a method to strongly increase the observable area by optically compensating for the meniscus effect. The microscope’s condenser annulus is replaced by a transmissive LCD to allow dynamic changes. A deformable, liquid-filled prism is placed in the illumination path. The prism’s surface angle is adaptively inclined to refract transmitted light so that the tangential angle of the liquid lens can be compensated. Besides the observation of the phase contrast image, a beam splitter allows to simultaneously view condenser annulus and phase ring displacement. Algorithms analyze the displacement to dynamically adjust the LCD and prism to guarantee phase contrast conditions. Experiments show a significant increase in observable area, especially for small well sizes. For 96-well-plates, more than twelve times the area can be examined under phase contrast conditions instead of standard phase contrast microscopy. Nature Publishing Group UK 2023-04-08 /pmc/articles/PMC10082838/ /pubmed/37031241 http://dx.doi.org/10.1038/s41598-023-32917-6 Text en © The Author(s) 2023, corrected publication 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Nienhaus, Florian Piotrowski, Tobias Nießing, Bastian König, Niels Schmitt, Robert H. Adaptive phase contrast microscopy to compensate for the meniscus effect |
title | Adaptive phase contrast microscopy to compensate for the meniscus effect |
title_full | Adaptive phase contrast microscopy to compensate for the meniscus effect |
title_fullStr | Adaptive phase contrast microscopy to compensate for the meniscus effect |
title_full_unstemmed | Adaptive phase contrast microscopy to compensate for the meniscus effect |
title_short | Adaptive phase contrast microscopy to compensate for the meniscus effect |
title_sort | adaptive phase contrast microscopy to compensate for the meniscus effect |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10082838/ https://www.ncbi.nlm.nih.gov/pubmed/37031241 http://dx.doi.org/10.1038/s41598-023-32917-6 |
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