Comparative Analysis of Electron Microscopy Techniques for Hydrogel Microarchitecture Characterization: SEM, Cryo-SEM, ESEM, and TEM

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Hydrogels have emerged as a versatile class of materials with broad applications in biomedical engineering, drug delivery, and tissue engineering. Understanding their intricate structures and morphologies is crucial for tailoring their properties to meet specific biomedical needs. It has been clearly established that the composition and microarchitecture of the materials play a critical role in essential cellular mechanisms such as mechanosensing, adhesion, and remodeling. This question is essential in tissue engineering, where precisely characterizing the microarchitecture of the materials used to model the cell microenvironment is a critical step to ensure the reproducibility and relevance of reconstructed tissues. In this study, we present a comprehensive comparison of four advanced electron microscopy techniques, namely, scanning electron microscopy, cryo-scanning electron microscopy, environmental scanning electron microscopy, and transmission electron microscopy, to observe the hydrogel microarchitecture, including a comparison of the sample preparation methods for each technique. Each technique's specific advantages and limitations are discussed in detail, highlighting their unique capabilities in characterizing the hydrogel structures. We illustrate this study with two semisynthetic hydrogels, such as gelatin methacrylate and hyaluronic acid methacrylate. Moreover, we delve into the critical sample preparation steps necessary for each method, emphasizing the need to preserve the hydrogel's native state while obtaining high-resolution images. This comparative analysis aims to select the most suitable electron microscopy technique for their hydrogel studies, fostering deeper insights into the design and development of advanced biomaterials for tissue engineering applications.

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