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Epifluorescence-based three-dimensional traction force microscopy

We introduce a novel method to compute three-dimensional (3D) displacements and both in-plane and out-of-plane tractions on nominally planar transparent materials using standard epifluorescence microscopy. Despite the importance of out-of-plane components to fully understanding cell behavior, epiflu...

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Autores principales: Hazlett, Lauren, Landauer, Alexander K., Patel, Mohak, Witt, Hadley A., Yang, Jin, Reichner, Jonathan S., Franck, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538907/
https://www.ncbi.nlm.nih.gov/pubmed/33024138
http://dx.doi.org/10.1038/s41598-020-72931-6
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author Hazlett, Lauren
Landauer, Alexander K.
Patel, Mohak
Witt, Hadley A.
Yang, Jin
Reichner, Jonathan S.
Franck, Christian
author_facet Hazlett, Lauren
Landauer, Alexander K.
Patel, Mohak
Witt, Hadley A.
Yang, Jin
Reichner, Jonathan S.
Franck, Christian
author_sort Hazlett, Lauren
collection PubMed
description We introduce a novel method to compute three-dimensional (3D) displacements and both in-plane and out-of-plane tractions on nominally planar transparent materials using standard epifluorescence microscopy. Despite the importance of out-of-plane components to fully understanding cell behavior, epifluorescence images are generally not used for 3D traction force microscopy (TFM) experiments due to limitations in spatial resolution and measuring out-of-plane motion. To extend an epifluorescence-based technique to 3D, we employ a topology-based single particle tracking algorithm to reconstruct high spatial-frequency 3D motion fields from densely seeded single-particle layer images. Using an open-source finite element (FE) based solver, we then compute the 3D full-field stress and strain and surface traction fields. We demonstrate this technique by measuring tractions generated by both single human neutrophils and multicellular monolayers of Madin–Darby canine kidney cells, highlighting its acuity in reconstructing both individual and collective cellular tractions. In summary, this represents a new, easily accessible method for calculating fully three-dimensional displacement and 3D surface tractions at high spatial frequency from epifluorescence images. We released and support the complete technique as a free and open-source code package.
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spelling pubmed-75389072020-10-07 Epifluorescence-based three-dimensional traction force microscopy Hazlett, Lauren Landauer, Alexander K. Patel, Mohak Witt, Hadley A. Yang, Jin Reichner, Jonathan S. Franck, Christian Sci Rep Article We introduce a novel method to compute three-dimensional (3D) displacements and both in-plane and out-of-plane tractions on nominally planar transparent materials using standard epifluorescence microscopy. Despite the importance of out-of-plane components to fully understanding cell behavior, epifluorescence images are generally not used for 3D traction force microscopy (TFM) experiments due to limitations in spatial resolution and measuring out-of-plane motion. To extend an epifluorescence-based technique to 3D, we employ a topology-based single particle tracking algorithm to reconstruct high spatial-frequency 3D motion fields from densely seeded single-particle layer images. Using an open-source finite element (FE) based solver, we then compute the 3D full-field stress and strain and surface traction fields. We demonstrate this technique by measuring tractions generated by both single human neutrophils and multicellular monolayers of Madin–Darby canine kidney cells, highlighting its acuity in reconstructing both individual and collective cellular tractions. In summary, this represents a new, easily accessible method for calculating fully three-dimensional displacement and 3D surface tractions at high spatial frequency from epifluorescence images. We released and support the complete technique as a free and open-source code package. Nature Publishing Group UK 2020-10-06 /pmc/articles/PMC7538907/ /pubmed/33024138 http://dx.doi.org/10.1038/s41598-020-72931-6 Text en © The Author(s) 2020 Open AccessThis 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/.
spellingShingle Article
Hazlett, Lauren
Landauer, Alexander K.
Patel, Mohak
Witt, Hadley A.
Yang, Jin
Reichner, Jonathan S.
Franck, Christian
Epifluorescence-based three-dimensional traction force microscopy
title Epifluorescence-based three-dimensional traction force microscopy
title_full Epifluorescence-based three-dimensional traction force microscopy
title_fullStr Epifluorescence-based three-dimensional traction force microscopy
title_full_unstemmed Epifluorescence-based three-dimensional traction force microscopy
title_short Epifluorescence-based three-dimensional traction force microscopy
title_sort epifluorescence-based three-dimensional traction force microscopy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538907/
https://www.ncbi.nlm.nih.gov/pubmed/33024138
http://dx.doi.org/10.1038/s41598-020-72931-6
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