Measuring Elastic Moduli of Human Trabecular Meshwork Tissues, Cells, or Extracellular Matrices

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Exploration of the mechanical properties of biological tissues has sparked a growing interest in the mechanobiology of various ocular diseases. There is an increasing recognition that the change in mechanical attributes (of cells/tissues) may reflect disease states or may be a viable endpoint to assess cell/tissue function. As such, mechanobiological investigations often rely on developing substrates with properties mimicking the mechanical properties of the choice of cell/tissue, either in homeostasis or in disease. Variability in methods of mechanical characterization has resulted in confounding reports that subsequently impact reproducibility in science and the development of appropriate scientific hypotheses. Here, we describe a protocol for measuring the elastic moduli of cells, matrices, and tissues relevant to the outflow pathway. We discuss factors to be considered while selecting cantilevers and preparing samples for measurements and provide detailed instructions for calibration of cantilevers, collection of force-displacement curves, and a workflow for how to analyze the data and determine elastic modulus. Although the protocol is intended for the cells/tissues of the iridocorneal angle, readers may apply or adapt this protocol for other relevant tissues.