Experimental investigation of effects of indentation depth and probe tip radius on apparent young's modulus of Xenopus laevis oocytes

Abstract

Throughout their lifespan cells are constantly subjected to mechanical forces. Numerous studies have been conducted to measure mechanical properties of animal cells and understand their effects of the cells’ biological processes. However, the range of the apparent Young's moduli reported in the literature spans three orders of magnitude from 0.1 kPa to 100 kPa, and no clear explanation is given for the source of variation. One of the commonly used experimental techniques to measure material properties at nano- and microscale is nanoindentation. Nanoindentation works by applying a force on a sample via a small probe and recording a force-displacement curve. Probes with different tip shapes and sizes are available. However, there is little guidance on the appropriate choice of probe geometry and size for measurement of properties of animal cells. Eukaryotic cells have a complex structure including a membrane, an actin cortex, a cytoplasm, a nucleus and a cytoskeleton which consists of microfilaments, intermediate filaments and microtubules. These components have different mechanical properties contributing to the overall properties of cells. This work focuses on the effect of tip size and indentation depth on the apparent Young's modulus of Xenopus laevis oocytes measuring approximately 1 mm in diameter. Live cell fluorescent labeling using Lifeact-EGFP and confocal microscopy was used to measure the thickness of the oocyte actin cortex which is expected to have a significant contribution to the cell's stiffness. Apparent mechanical properties were measured using the JPK NanoWizard 4 BioAFM system. Indentations 5, 100 and 400 nm deep using probes with spherical tips of 0.1, 1, and 10 μm radii were performed. Contributions of the membrane and the actin cortex on the overall mechanical properties of the oocytes are analyzed.