Three-Dimensional Reflectance Traction Microscopy

Kim, Jihan; Jones, Christopher; Groves, Nicholas; Sun, Bo

doi:10.1371/journal.pone.0156797

Cited by 22 publications

(29 citation statements)

References 49 publications

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“…One solution to this problem may be to abandon scattering beads entirely. TFM has been demonstrated using confocal reflectance microscopy to track the motion of collagen fibers, instead of embedded beads 35 . OCT/OCM could similarly be used to image the deformation of fibrous extracellular matrix constituents for TF-OCM.…”

Section: Discussionmentioning

confidence: 99%

Quantitative reconstruction of time-varying 3D cell forces with traction force optical coherence microscopy

Mulligan

Feng

Adie

2018

Preprint

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11Cellular traction forces (CTFs) play an integral role in both physiological processes and disease, and are a topic of 12 interest in mechanobiology. Traction force microscopy (TFM) is a family of methods used to quantify CTFs in a 13 variety of settings. State-of-the-art 3D TFM methods typically rely on confocal fluorescence microscopy, which can 14 impose limitations on acquisition speed, volumetric coverage, and temporal sampling or coverage. In this report, we 15 present the first quantitative implementation of a new TFM technique: traction force optical coherence microscopy 16(TF-OCM). TF-OCM leverages the capabilities of optical coherence microscopy and computational adaptive optics 17(CAO) to enable the quantitative reconstruction of 3D CTFs in scattering media with minute-scale temporal 18 resolution. We applied TF-OCM to quantify CTFs exerted by isolated NIH-3T3 fibroblasts embedded in Matrigel, 19with five-minute temporal sampling, using images which spanned a 500×500×500 μm 3 field-of-view. Due to the 20 reliance of TF-OCM on computational imaging methods, we have provided extensive discussion of the underlying 21 equations, assumptions, and failure modes of these methods. TF-OCM has the potential to advance studies of 22 biomechanical behavior in scattering media, and may be especially well-suited to the study of cell collectives such 23as spheroids, a prevalent model in mechanobiology research. 24

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Section: Discussionmentioning

confidence: 99%

Quantitative reconstruction of time-varying 3D cell forces with traction force optical coherence microscopy

Mulligan

Feng

Adie

2018

Preprint

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“…In principal, TFM can be adapted to 3D (Hall et al, 2013). 3D ECM deformations can be measured using confocal reflectance or optical coherence microscopy (Kim et al, 2016;Mulligan et al, 2017) in addition to using tracer beads ( Figure 1F). However, accurate 3D stress mapping is feasible mostly in synthetic materials that are linearly elastic, homogenous and isotropic, such as polyethylene glycol (PEG) hydrogels (Legant et al, 2010).…”

Section: Traction Force Microscopy (Tfm)mentioning

confidence: 99%

Microscale Interrogation of 3D Tissue Mechanics

Zhang

Chada

Reinhart‐King

2019

Front. Bioeng. Biotechnol.

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Cells in vivo live in a complex microenvironment composed of the extracellular matrix (ECM) and other cells. Growing evidence suggests that the mechanical interaction between the cells and their microenvironment is of critical importance to their behaviors under both normal and diseased conditions, such as migration, differentiation, and proliferation. The study of tissue mechanics in the past two decades, including the assessment of both mechanical properties and mechanical stresses of the extracellular microenvironment, has greatly enriched our knowledge about how cells interact with their mechanical environment. Tissue mechanical properties are often heterogeneous and sometimes anisotropic, which makes them difficult to obtain from macroscale bulk measurements. Mechanical stresses were first measured for cells cultured on two-dimensional (2D) surfaces with well-defined mechanical properties. While 2D measurements are relatively straightforward and efficient, and they have provided us with valuable knowledge on cell-ECM interactions, that knowledge may not be directly applicable to in vivo systems. Hence, the measurement of tissue stresses in a more physiologically relevant three-dimensional (3D) environment is required. In this mini review, we will summarize and discuss recent developments in using optical, magnetic, genetic, and mechanical approaches to interrogate 3D tissue stresses and mechanical properties at the microscale.

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“…A recent alternative for measuring substratum deformation involves imaging deformations of fibrous matrix materials as opposed to embedded beads [141,142]. These studies offer a new approach for calculating 3D displacement fields that may be advantageous in systems where bead displacement techniques are not feasible or have limited accuracy.…”

Section: Strategies For Overcoming the Limitations Of Conventional Tfmmentioning

confidence: 99%

“…These studies offer a new approach for calculating 3D displacement fields that may be advantageous in systems where bead displacement techniques are not feasible or have limited accuracy. For example, during remodeling of a fibrous matrix, reflection microscopy can be used to calculate matrix displacements and obtain qualitative information on cell tractions [141,142]. Quantitative cell traction calculations rely on the assumption that the fibrous network is isotropic and can be approximated as linearly elastic [141].…”

Section: Strategies For Overcoming the Limitations Of Conventional Tfmmentioning

confidence: 99%

“…For example, during remodeling of a fibrous matrix, reflection microscopy can be used to calculate matrix displacements and obtain qualitative information on cell tractions [141,142]. Quantitative cell traction calculations rely on the assumption that the fibrous network is isotropic and can be approximated as linearly elastic [141]. In addition, molecular force sensors offer one strategy for improving the spatial resolution of quantified traction forces [143–147].…”

Section: Strategies For Overcoming the Limitations Of Conventional Tfmmentioning

confidence: 99%

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Microfabricated tissues for investigating traction forces involved in cell migration and tissue morphogenesis

Nerger

Siedlik

Nelson

2016

Cell. Mol. Life Sci.

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Cell-generated forces drive an array of biological processes ranging from wound healing to tumor metastasis. Whereas experimental techniques such as traction force microscopy are capable of quantifying traction forces in multidimensional systems, the physical mechanisms by which these forces induce changes in tissue form remain to be elucidated. Understanding these mechanisms will ultimately require techniques that are capable of quantifying traction forces with high precision and accuracy in vivo or in systems that recapitulate in vivo conditions, such as microfabricated tissues and engineered substrata. To that end, here we review the fundamentals of traction forces, their quantification, and the use of microfabricated tissues designed to study these forces during cell migration and tissue morphogenesis. We emphasize the differences between traction forces in two- and three-dimensional systems, and highlight recently developed techniques for quantifying traction forces.

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Three-Dimensional Reflectance Traction Microscopy

Cited by 22 publications

References 49 publications

Quantitative reconstruction of time-varying 3D cell forces with traction force optical coherence microscopy

Quantitative reconstruction of time-varying 3D cell forces with traction force optical coherence microscopy

Microscale Interrogation of 3D Tissue Mechanics

Microfabricated tissues for investigating traction forces involved in cell migration and tissue morphogenesis

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