Scanning electron microscopy of murine skin ultrathin sections and cultured keratinocytes

The document outlined a protocol for using scanning electron microscopy (SEM) to examine the ultrastructure of murine skin and cultured keratinocytes, focusing on details like the epidermis, dermis, hair follicles, and cell-cell junctions. It described the preparation of ultrathin sections and cell monolayers, including steps like fixation, dehydration, and embedding, with an emphasis on proper sample orientation. The study utilized both male and female mice, with newborn pups for cell monolayer protocols and 3-8 week old animals for skin samples. The protocol enabled visualization of skin structures such as hair follicles and sebaceous glands, as well as subcellular structures like cell junctions and mitochondria. Limitations of SEM, such as its resolution compared to TEM, were discussed, along with troubleshooting tips for common issues in sample preparation and imaging.