Background:In AFM study of cell mechanical properties, the apparent elastic modulus of a cell is affected by many factors, especially the AFM tip geometry, indentation site of the cell, the application of the mathematical model and testing conditions.Background:In AFM study of cell mechanical properties, the apparent elastic modulus of a cell is affected by many factors, especially the AFM tip geometry, indentation site of the cell, the application of the mathematical model and testing conditions.Methods:In this study, indentation experiments of living cells under different conditions were performed aiming to build an accurate evaluation system of mechanical properties of lung cancer cells based on AFM. Comparisons of the effects of spherical and pyramid AFM tips, Hertz model of semiinfinite and finite thickness, cell nuclear and cytoplasmic indentation regions on the cell apparent elastic modulus were accomplished.Methods:In this study, indentation experiments of living cells under different conditions were performed aiming to build an accurate evaluation system of mechanical properties of lung cancer cells based on AFM. Comparisons of the effects of spherical and pyramid AFM tips, Hertz model of semiinfinite and finite thickness, cell nuclear and cytoplasmic indentation regions on the cell apparent elastic modulus were accomplished.Results:Compared with the calculated results by spherical tip, the elastic modulus distribution of non-small lung cancer cells (NSCLC) by pyramid tip was observed to be similar while the absolute values increased obviously, which were more than twice the numerical values by the spherical tip (p<0.05). The apparent elastic modulus values were the overvalued cause of the underestimation of the contact region in pyramidal tip measurement. Two different indentations over nucleus or lamellipodium of NCI-H520 cell and NCI-H1299 cell were analyzed. Consequently, the exact elastic modulus over the nucleus area can be calculated accurately using the semi-infinite Hertz model while the finite thickness Hertz model should be used for elasticity assessment of cell lamellipodium with a small thickness.Results:Compared with the calculated results by spherical tip, the elastic modulus distribution of non-small lung cancer cells (NSCLC) by pyramid tip was observed to be similar while the absolute values increased obviously, which were more than twice the numerical values by the spherical tip (p<0.05). The apparent elastic modulus values were the overvalued cause of the underestimation of the contact region in pyramidal tip measurement. Two different indentations over nucleus or lamellipodium of NCI-H520 cell and NCI-H1299 cell were analyzed. Consequently, the exact elastic modulus over the nucleus area can be calculated accurately using the semi-infinite Hertz model while the finite thickness Hertz model should be used for elasticity assessment of cell lamellipodium with a small thickness.Conclusion:This evaluation system provides technological support for accurate evaluation of viscoelastic properties of living cancer cells.Conclusion:This evaluation system provides technological support for accurate evaluation of viscoelastic properties of living cancer cells.