Utilizing collagen-coated hydrogels with defined stiffness as calibration standards for AFM experiments on soft biological materials: the case of lung cells and tissue

• DOI: 10.1515/mim-2024-0014
• Publication Date: 2024-11-04T11:33:29Z
• Authors: Andreas Stylianou; Stylianos Vasilei...; Katerina Polemidi...; Andreas Zachariades

Abstract Atomic Force Microscopy (AFM) is crucial in mechanobiology for high-resolution imaging and nanomechanical measurements of biological samples, providing insights into their mechanical properties. However, AFM faces challenges such as tip damage and cantilever selection errors, impacting measurement accuracy. This study proposes a methodology using collagen-coated hydrogels with predefined stiffness for calibrating AFM measurements on soft biological materials. By facilitating appropriate cantilever selection, assessing systematic errors, and evaluating tip damage, this approach ensures reliable Young’s modulus measurements. The proof of concept with human lung cells and tissue specimens demonstrates improved accuracy and reliability of AFM-based nanomechanical characterizations, essential for understanding cellular mechanics and disease progression.