Measuring Cell Mechanics with AFM

Stiffness of cell nucleus and cytoplasm was measured with a NanoWizard 4 AFM (JPK) instrument mounted on top of a Nikon Ti ECLIPSE microscope^{62 (link)}. The spring constant of the cantilevers was calibrated by thermal tuning using the simple harmonic oscillator model. The Hertz model was fitted to the approach curves to obtain the stiffness value using the JPKSPM Data Processing software (version 6.1.79). The cells were seeded on laminin-111-coated coverslips and a force curve on top of the nucleus and cytoplasm was acquired for each of the cells. The cells were kept at 37 °C using a BioCell (JPK) and maintained in a CO₂-independent medium (Thermo Fisher, category no. 18045088).

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Kechagia Z., Sáez P., Gómez-González M., Canales B., Viswanadha S., Zamarbide M., Andreu I., Koorman T., Beedle A.E., Elosegui-Artola A., Derksen P.W., Trepat X., Arroyo M, & Roca-Cusachs P. (2023). The laminin–keratin link shields the nucleus from mechanical deformation and signalling. Nature Materials, 22(11), 1409-1420.

Publication 2023

NanoWizard 4 AFM Ti ECLIPSE microscope BioCell

Cells Cytoplasm Laminin Nucleus

Corresponding Organization : Institute for Bioengineering of Catalonia

Other organizations : Universitat Politècnica de Catalunya, The Francis Crick Institute, Universitat de Barcelona, University Medical Center Utrecht

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Stiffness of cell nucleus
Stiffness of cytoplasm

control variables

Cell seeding on laminin-111-coated coverslips
Maintaining cells at 37 °C using a BioCell (JPK)
Maintaining cells in a CO2-independent medium (Thermo Fisher, category no. 18045088)

controls

Positive control: None mentioned
Negative control: None mentioned

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