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Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy
In spite of the advantages associated with the molecular specificity of fluorescence imaging, there is still a significant need to augment these approaches with label-free imaging. Therefore, we have implemented a form of interference microscopy based upon phase-shifted, laser-feedback interferometr...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Optical Society of America
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3149539/ https://www.ncbi.nlm.nih.gov/pubmed/21833378 http://dx.doi.org/10.1364/BOE.2.002417 |
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author | Atılgan, Erdinç Ovryn, Ben |
author_facet | Atılgan, Erdinç Ovryn, Ben |
author_sort | Atılgan, Erdinç |
collection | PubMed |
description | In spite of the advantages associated with the molecular specificity of fluorescence imaging, there is still a significant need to augment these approaches with label-free imaging. Therefore, we have implemented a form of interference microscopy based upon phase-shifted, laser-feedback interferometry and developed an algorithm that can be used to separate the contribution of the elastically scattered light by sub-cellular structures from the reflection at the coverslip-buffer interface. The method offers an opportunity to probe protein aggregation, index of refraction variations and structure. We measure the topography and reflection from calibration spheres and from stress fibers and adhesions in both fixed and motile cells. Unlike the data acquired with reflection interference contrast microscopy, where the reflection from adhesions can appear dark, our approach demonstrates that these regions have high reflectivity. The data acquired from fixed and live cells show the presence of a dense actin layer located ≈ 100 nm above the coverslip interface. Finally, the measured dynamics of filopodia and the lamella in a live cell supports retrograde flow as the dominate mechanism responsible for filopodia retraction. |
format | Online Article Text |
id | pubmed-3149539 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Optical Society of America |
record_format | MEDLINE/PubMed |
spelling | pubmed-31495392011-08-10 Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy Atılgan, Erdinç Ovryn, Ben Biomed Opt Express Cell Studies In spite of the advantages associated with the molecular specificity of fluorescence imaging, there is still a significant need to augment these approaches with label-free imaging. Therefore, we have implemented a form of interference microscopy based upon phase-shifted, laser-feedback interferometry and developed an algorithm that can be used to separate the contribution of the elastically scattered light by sub-cellular structures from the reflection at the coverslip-buffer interface. The method offers an opportunity to probe protein aggregation, index of refraction variations and structure. We measure the topography and reflection from calibration spheres and from stress fibers and adhesions in both fixed and motile cells. Unlike the data acquired with reflection interference contrast microscopy, where the reflection from adhesions can appear dark, our approach demonstrates that these regions have high reflectivity. The data acquired from fixed and live cells show the presence of a dense actin layer located ≈ 100 nm above the coverslip interface. Finally, the measured dynamics of filopodia and the lamella in a live cell supports retrograde flow as the dominate mechanism responsible for filopodia retraction. Optical Society of America 2011-07-27 /pmc/articles/PMC3149539/ /pubmed/21833378 http://dx.doi.org/10.1364/BOE.2.002417 Text en ©2011 Optical Society of America http://creativecommons.org/licenses/by-nc-nd/3.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 Unported License, which permits download and redistribution, provided that the original work is properly cited. This license restricts the article from being modified or used commercially. |
spellingShingle | Cell Studies Atılgan, Erdinç Ovryn, Ben Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy |
title | Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy |
title_full | Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy |
title_fullStr | Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy |
title_full_unstemmed | Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy |
title_short | Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy |
title_sort | reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy |
topic | Cell Studies |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3149539/ https://www.ncbi.nlm.nih.gov/pubmed/21833378 http://dx.doi.org/10.1364/BOE.2.002417 |
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