Abstract

An important factor affecting the accuracy of Young's modulus calculation in Atomic Force Microscopy (AFM) indentation experiments is the determination of the dimensions of the indenter. This procedure is usually performed using AFM calibration gratings or Scanning Electron Microscopy (SEM) imaging. However, the aforementioned procedure is frequently omitted because it requires additional equipment. In this paper, a new approach is presented that focused on the calibration of spherical indenters without the need of special equipment but instead using force indentation data on soft samples. Firstly, the question whether it is mathematically possible to simultaneously calculate the indenter's radius and the Young's modulus of the tested sample (under the restriction that the sample presents a linear elastic response) using the same force indentation data is discussed. Using a simple mathematical approach, it was proved that the aforementioned procedure is theoretically valid. In addition, to test this method in real indentation experiments agarose gels were used. Multiple measurements on different agarose gels showed that the calibration of a spherical indenter is possible and can be accurately performed. Thus, the indenter's radius and the soft sample's Young's modulus can be determined using the same force indentation data. It is also important to note that the provided accuracy is similar to the accuracy obtained when using AFM calibration gratings. The major advantage of this paper is that it provides a method for the simultaneous determination of the indenter's radius and the sample's Young's modulus without requiring any additional equipment.

Highlights

  • Spherical indenters are frequently used in Atomic Force Microscopy (AFM) indentation experiments on soft biological samples [1–6]

  • According to the AtomicJ repository, the simulated curves were generated in Mathematica 8.0 as if they were real force curves, using a spherical indenter

  • A method for calibrating spherical indenters used in AFM indentation experiments regarding soft biological samples was presented and discussed

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Summary

Introduction

Spherical indenters are frequently used in Atomic Force Microscopy (AFM) indentation experiments on soft biological samples [1–6]. They are preferable since they usually do not cause permanent damage to the abovementioned samples during indentation (at least in cases that the indentation depth is not significantly bigger compared to the indenter’s radius). In equation (1), E and v are the material’s Young’s modulus and Poisson’s ratio, respectively, and R is the sphere’s radius. The Young’s modulus of the sample of interest can be accurately determined under the condition that the sample’s Poisson’s ratio and the indenter’s radius are known. It is significant to determine the indenter’s radius in order to accurately calculate the sample’s Young’s modulus.

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