Fig. 1

Schematic view of the procedure for whole-mount in situ hybridization optimized for mouse oocytes and embryos. A The structure of glass pipettes used in the procedure. B After the oocytes/embryos had been collected in a 24-well cell culture plate, the samples were fixed with 4% PFA/PBS, permeabilized with 2% Triton X-100, and incubated with 66% formamide/10% SSC (pre-hybridization). A 24-well cell culture plate was used in these steps. Then the oocytes/embryos were transferred to a 96-well cell culture plate and hybridized with probes of target RNAs at 45˚C overnight. The samples were washed with a series of SSC and incubated with blocking buffer. Then anti-DIG or Fluorescein antibody conjugated with horseradish peroxidase (HRP) was added, and signals were amplified with the TSA system. A 96-well cell culture plate was used in these steps. Finally, the samples were mounted on a slide glass with mounting medium with DAPI