Three-dimensional label-free imaging and quantification of migrating cells during wound healing

Abstract

The wound-healing assay is a simple but effective tool for studying collective cell migration (CCM) that is widely used in biophysical studies and high-throughput screening. However, conventional imaging and analysis methods only address two-dimensional (2D) properties in a wound healing assay, such as gap closure rate. This is unfortunate because biological cells are complex 3D structures, and their dynamics provide significant information about cell physiology. Here, we presented 3D label-free imaging for wound healing assays and investigated the 3D dynamics of CCM using optical diffraction tomography. High-resolution subcellular structures as well as their collective dynamics were imaged and analyzed quantitatively.

Quantitative Phase Imaging: Recent Advances and Expanding Potential in Biomedicine

“…Measurements of cell mass and morphology with the same system could similarly track kinetic epithelial-to-mesenchymal cell transitions in heterogeneous cultures . Finally, optical diffraction tomography, a 3D, label-free QPI-based imaging method, was used to study and quantify the dynamics of NIH3T3 cell migration in a wound healing assay, revealing single cell resolution of subcellular structure behavior and transport that underlies the mechanisms involved in gap closure and closure rate, with potential implications for pharmaceuticals development or repurposing …”

“…234 Finally, optical diffraction tomography, a 3D, label-free QPIbased imaging method, was used to study and quantify the dynamics of NIH3T3 cell migration in a wound healing assay, revealing single cell resolution of subcellular structure behavior and transport that underlies the mechanisms involved in gap closure and closure rate, with potential implications for pharmaceuticals development or repurposing. 235 Applications of QPI for Measuring Biophysical Cell Properties. QPI can measure the distribution of mass within a cell, including mass due to structural elements such as the cytoskeleton, and how this distribution changes over time.…”

Computational Investigation of Cell Migration Behavior in a Confluent Epithelial Monolayer

“…The epithelial monolayer is reported to be a thin layer of 3-15 μm in thickness [43,44]. For simplification, a 2D plane stress model of the epithelial monolayer was created using the Voronoi tessellation method to represent the monolayer sheet [35,45,46].…”

Advances in Digital Holographic Interferometry

“…In conclusion of this review, we would like to note the most impressive achievement related to digital holographic interferometry. The combination of digital holography phase-contrast microscopy methods led to the creation of a unique technology—holographic tomography [ 4 , 87 , 88 , 89 ]. This technology allows you to simultaneously study the thickness of the object (absorption) and the refractive index.…”