Sd sphere cont m 10

An AFM (Keysight 5500, Keysight Technologies, Santa Rosa, CA, USA) was used to analyze 20 μm thick fresh frozen tissue sections without decalcification. For the micromorphological imaging, after the sections were washed with water to remove the embedding medium (O.C.T compound) and dried naturally. A silicon probe (PPP-NCLR-20, Nanosensors™, Neuchatel, Switzerland) with a force constant of 42 N/m and a resonance frequency of 161 kHz was used for imaging. For measurement of mechanical properties, the AFM-based nanoindentation was used. The washed specimens were immersed in physiological saline at room temperature, and a silicon oxide tip with 13 kHz resonance frequency and 0.2 N/m force constant (SD-Sphere-CONT-M-10, Nanosensors) was used in the contact mode. The elastic modulus of each position was measured at an indentation depth of 300 nm. All measurements were repeated for six positions in each tissue specimen, and the values were averaged.