Holistic approach to visualize and quantify collagen organization at macro, micro, and nano‐scale

BACKGROUND: There is scarcity of imaging and image processing techniques for accurate discrimination and quantitation of the dermal extracellular matrix (ECM), primarily collagen. The aim of this study was to develop and demonstrate a holistic imaging and image processing approach to visualize and quantify collagen remodeling at the macro‐, micro‐ and nano‐scale using histochemical imaging, Reflectance Confocal Microscopy (RCM), and Atomic Force Microscopy (AFM), respectively. MATERIAL AND METHODS: For proof‐of‐concept, a commercial anti‐aging product known to induce collagen neo‐synthesis and re‐organization was tested ex vivo on human skin biopsies from two aged females. RESULTS: Relative to untreated skin, collagen fibers (RCM) and fibrils (AFM) were longer and aligned after treatment. The content of collagen and elastin (histochemical imaging and ELISA) statistically improved after treatment. CONCLUSION: Based on our findings, we can conclude: (1) AFM, RCM, and histochemical imaging can accurately discriminate collagen from other ECM components in the skin and (2) the image processing methods can enable quantitation and hence capture small improvements in collagen remodeling after treatment (commercial cosmetic product with collagen organizer technology as proof‐of‐concept). The reported holistic imaging approach has direct clinical implications for scientists and dermatologists to make quick, real‐time, and accurate decisions in skin research and diagnostics.