Ex Vivo SIM-AFM Measurements Reveal the Spatial Correlation of Stiffness and Molecular Distributions in 3D Living Tissue

    October 2024 in “ Acta Biomaterialia ”
    Itsuki Shioka, Ritsuko Morita, Rei Yagasaki, Duligengaowa Wuergezhen, Tadahiro Yamashita, Hironobu Fujiwara, Satoru Okuda
    TLDR Collagen makes skin stiff, and preservation methods greatly increase tissue stiffness.
    The study introduces a novel technique, USIM-AFM, which combines atomic force microscopy (AFM) and structured illumination microscopy (SIM) to map stiffness and molecular distributions in 3D living tissues, using mouse embryonic and adult skin as models. It reveals that collagen significantly contributes to skin stiffness, with enzymatic digestion reducing stiffness in both epithelium and mesenchyme. The study finds that preservation processes like freeze-thaw, PFA, and glyoxal fixation significantly alter tissue stiffness, increasing it by about 10-fold, although the relative stiffness between epithelium and mesenchyme is maintained. The USIM-AFM technique provides high-resolution stiffness maps and fluorescence images, offering insights into the mechanical properties of tissues and highlighting the importance of live measurements for accurate tissue stiffness characterization.
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