Deformation Microscopy: A Protocol for Full-Field, Micro- to Nano-Scale Imaging of Mechanics in Extracellular Matrix, Cells, and Nuclei. Journal Article

Overview

The internal mechanics of living tissues, cells, and nuclei exhibit exquisite complexity, depend on dynamic structural changes, and influence biological functions such as tissue remodeling, cell homeostasis and migration, and gene expression. Full-field methods to reveal the internal mechanics of extracellular matrix, cells, and nuclei are emerging and can uncover new discoveries in mechanobiology. Here we detail a noninvasive technology to probe the internal mechanics of extracellular matrix, cells, and nuclei using a combination of imaging and deformation-matching computational approaches. Important to this technology is the acquisition of image data showing sufficient spatial and temporal resolution to capture a motion event, which can be readily evaluated using automated assessment of structural features. With careful consideration of image acquisition and processing parameters, micron- to nanometer-scale intracellular and intranuclear mechanics can be reliably measured with small displacement errors (<0.04 µm) using a conventional confocal or wide-field microscope. Further, the technology can be used to spatially correlate a multitude of biological events in complex biological problems. © 2026 Wiley Periodicals LLC. Basic Protocol: Quantifying cellular and nuclear deformation using deformation microscopy.