After lyophilization, while scoping out the material it affects its structure.
Yes, there are other techniques for preparing hydrogel samples for SEM analysis apart from lyophilization. One approach is **dehydration using a gradation series of ethanol**, followed by an additional drying step such as air drying, drying in hexamethyldisilazane, critical point drying or freeze-drying ¹. Another approach is **freeze-drying of fresh, water-containing samples** ¹. There is also a **high pressure water freeze fracture method** for preparing SEM samples from hydrogels without using organic solvents ².
(1) Faithful Scanning Electron Microscopic (SEM) visualization of 3D .... https://www.biorxiv.org/content/biorxiv/early/2020/03/20/2020.03.18.997668.full.pdf.
(2) Preparation of SEM hydrogel samples using a high pressure water freeze .... https://www.tandfonline.com/doi/full/10.1080/08957959.2020.1863399.
(3) Scanning Electron Microscope | SEM Sample Preparation | Thermo ... - US. https://www.thermofisher.com/us/en/home/materials-science/learning-center/applications/sample-preparation-techniques-sem.html.
(4) Morphological Characterization of Hydrogels | SpringerLink. https://link.springer.com/referenceworkentry/10.1007/978-3-319-77830-3_28.
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