Axio m1 microscope

Melanoma cells were treated with PXDN, NTN4 or GLIS3-specific siRNAs or scrambled control siRNA as described above and labelled with CM-DiO or Dil as per manufacturer's instructions (Invitrogen, USA). Cells were grown overnight in a hanging-drop fashion to allow the formation of aggregates. Fertile chicken eggs were incubated at 38°C for 48 hours prior to transplantation. Cell aggregates consisting of similar sized small aggregates from approximately 500–800 cells were harvested and carefully injected with a glass pipette into the trunk neural tube lumen of developing chicken embryos. The eggs were then sealed with adhesive tape and re-incubated for 2 days. After incubation, embryos were removed from the eggs and fixed with 4% paraformaldehyde and whole mounts were analyzed for the localization of melanoma cells using Lumar V12 Zeiss microscope as described previously [20 (link)]. Segments of embryos containing the melanoma cells were then washed in PBS, incubated overnight in 30% sucrose at 4°C, frozen and 20μm transverse sections cut using a cryostat. Sections were rinsed in PBS, mounted in fluorescent mounting media and imaged using a Zeiss Axio M1 microscope and AxioVision software.

To examine the expression of Mex3c in the mammary gland through detecting β-galactosidase (β-gal) activity, the fourth mammary glands of wildtype or heterozygous Mex3c gene trap mice were dissected and stained with X-Gal as described [22 (link)]. Some mammary glands stained with X-Gal were then stained with Carmine alum to visualize the mammary gland epithelium. Stained tissues were then embedded in paraffin and sectioned to observe under the microscope. Images were taken with an Axio M1 microscope equipped with an AxioCam MRc digital camera (Carl Zeiss).