2.2.3. Hair Follicle Preparation for Immunohistochemistry

The same process described for HF preparation for infrared analysis was used to prepare sections. Sections were placed on glass slides and air-dried; then, they were fixed in acetone for 10 min at –20 ◦C. After three washes in PBS, the sections were placed in a sheep serum solution (Thermo Fisher Scientific, Illkirch-Graffenstaden, France). Primary antibody anti-GPC1 (Proteintech, Rosemont, IL, USA) was incubated overnight at 4 ◦C. After several washes with PBS, the secondary antibody coupled with Alexa 488 was applied for 45 min at room temperature and in the dark. The Evans blue counterstain was applied after several washes for 5 min at room temperature. After the final washes, the glass slides were mounted under a coverslip using Fluoprep. The observations were performed using a confocal microscope (TCS-SPE, Leica Biosystems).
