Slide scannerz1

Hairs were plucked at the end of the first and second cycles (P20 and P50, respectively). For hair type analysis, hair shafts were placed on a two-sided adhesive tape slides and were counted under the Zeiss stereomicroscope (Discovery.V12). The analysis was performed on both mutant and control mice (n = 5 each), and at least 200 hairs were scored per mouse. To photograph hair shafts, plucked hairs were randomly mounted on slides in a thin layer of Gelvatol and were imaged in bright field with Zeiss upright AxioImagerM2 through a 20X objective. For hair length analysis, hair shafts were placed on a two-sided adhesive tape slides and imaged with Zeiss slide scannerZ1 through a 20x objective. The length was measured with Carl Zeiss’ image analysis tool for Zen Blue edition. 5 mice of each genotype were used, and ten hairs for each hair type were measured per mouse.

Imaging of in situ hybridization and X-gal staining was performed with Zeiss upright AxioImagerM2 through a 20× objective with tiling mode using Zen Blue 2.3 software. HE staining was imaged with Zeiss upright AxioImagerM2 or Zeiss slide scannerZ1 through a 20× objective. For immunofluorescence, Zeiss LSM780 inverted confocal microscope was used to acquire images through a 20× objective using the Zen Black 11 (service pack 7) software. Adobe Photoshop CS5.1 was employed to process all images.