We present a procedure that allows a reliable determination of the elastic (Young's) modulus of soft samples, including living cells, by atomic force microscopy (AFM). The standardized nanomechanical AFM procedure (SNAP) ensures the precise adjustment of the AFM optical lever system, a prerequisite for all kinds of force spectroscopy methods, to obtain reliable values independent of the instrument, laboratory and operator. Measurements of soft hydrogel samples with a well-defined elastic modulus using different AFMs revealed that the uncertainties in the determination of the deflection sensitivity and subsequently cantilever's spring constant were the main sources of error. SNAP eliminates those errors by calculating the correct deflection sensitivity based on spring constants determined with a vibrometer. The procedure was validated within a large network of European laboratories by measuring the elastic properties of gels and living cells, showing that its application reduces the variability in elastic moduli of hydrogels down to 1%, and increased the consistency of living cells elasticity measurements by a factor of two. The high reproducibility of elasticity measurements provided by SNAP could improve significantly the applicability of cell mechanics as a quantitative marker to discriminate between cell types and conditions.
Standardized Nanomechanical Atomic Force Microscopy Procedure (SNAP) for Measuring Soft and Biological Samples / H. Schillers, C. Rianna, J. Schäpe, T. Luque, H. Doschke, M. Wälte, J.J. Uriarte, N. Campillo, G.P.A. Michanetzis, J. Bobrowska, A. Dumitru, E.T. Herruzo, S. Bovio, P. Parot, M. Galluzzi, A. Podestà, L. Puricelli, S. Scheuring, Y. Missirlis, R. Garcia, M. Odorico, J. Teulon, F. Lafont, M. Lekka, F. Rico, A. Rigato, J. Pellequer, H. Oberleithner, D. Navajas, M. Radmacher. - In: SCIENTIFIC REPORTS. - ISSN 2045-2322. - 7:1(2017 Jul 11), pp. 5117.1-5117.9.
|Titolo:||Standardized Nanomechanical Atomic Force Microscopy Procedure (SNAP) for Measuring Soft and Biological Samples|
|Parole Chiave:||Atomic Force Microscopy; cell mechanics; nano-mechanics; standardization; health and disease; mechanical phenotyping|
|Settore Scientifico Disciplinare:||Settore FIS/03 - Fisica della Materia|
Settore FIS/07 - Fisica Applicata(Beni Culturali, Ambientali, Biol.e Medicin)
|Data di pubblicazione:||11-lug-2017|
|Centro di ricerca:||Centro Interdisciplinare Materiali ed Interfacce Nanostrutturati - CIMAINA|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1038/s41598-017-05383-0|
|Appare nelle tipologie:||01 - Articolo su periodico|
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