SEM Sample Preparation: A Detailed Guide with Step-by-Step Instructions

The sample preparation process for SEM was conducted as follows: First, the samples were fixed with 2.5% glutaraldehyde and 1% osmic acid, one after the other. Next, the samples were dehydrated using a series of ethanol solutions with increasing concentrations (50%, 70%, 90%, 100%, 100%) and 100% acetone solution. After dehydration, the samples were embedded in resin and then sectioned using a Leica UC7. The sections were stained with uranyl acetate and lead citrate. Finally, the ultra-thin sections were analyzed using a Tecnai G2 spirit equipped with a camera, operating at 120kV (FEI, USA).