Epifluorescence-based three-dimensional traction force microscopy
File(s)
Date
2020Author
Hazlett, Lauren
Landauer, Alexander K.
Patel, Mohak
Witt, Hadley
Yang, Jin
Reichner, Jonathan
Franck, Christian
Metadata
Show full item recordAbstract
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 understand 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.
Subject
traction force microscopy
epifluorescence microscopy
finite element analysis
neutrophils
Madin-Darby canine kidney (MDCK) cells
collective cell forces
Permanent Link
http://digital.library.wisc.edu/1793/80302Description
The datasets presented here are intended to be used with the Single-Layer-3D-TFM Matlab and FEniCS code package on the Franck Lab Github page (https://github.com/FranckLab). The datasets include: an example dataset for new users to practice using the code along with the experimental rigid displacement data, synthetic traction validation cases, and experimental cell traction data which can be used to reproduce the figures in the publication.