“…While the snap freezing and sectioning required in this method may alter mechanical properties, causing differences as compared to the in vivo state, snap freezing allows for long-term tissue storage, more uniform and thin sectioning, and has been widely used in the biomedical research field (Graham et al, 2010;Peña et al, 2022;Usukura et al, 2017;Wang et al, 2017). Rapid freezing and thawing has been shown to preserve biomechanical properties of tissue sections, and a consistent experimental protocol still allows researchers to compare the effects of different biological conditions or sample locations on tissue mechanical properties (Boettcher et al, 2014;Calò et al, 2020;Lopez et al, 2011;Tran et al, 2017). AFM force-displacement measurements have thus been performed on cryosections of various tissue types, including brain, heart, lens, cornea, retina, trabecular meshwork/Schlemm's canal, and optic nerve (Franze et al, 2011;Last et al, 2010;Menal et al, 2018;Perea-Gil et al, 2015;Vahabikashi et al, 2019;Wang et al, 2018Wang et al, , 2017.…”