Fig5d Sperm staining of caudal epididymal sperms.

Raw image dataset for sperm staining of caudal epididymal sperms from Wdr62 genetrap and control mice.
An
incision was made to the caudal end of isolated epididymus in DMEM (Gibco).
This was followed by 15 min incubation at 37°C
to allow spermatozoa to swim out. Sperms
were centrifugated at 150g for 5 min to remove DMEM. Sperms
were fixed in 95% ethanol for 15 min and 10μL
was smeared onto a glass slide and air-dried for 2 hrs. Slides were kept at 4°C overnight. The next day, sperm stain was
performed using the Papanicolaou method as described in https://doi.org/10.1093/eshremonographs/2002.2.13,
with 3 min incubation in Harris hematoxylin (Sigma), 2 min in orange G6 (OG6
0.5% solution in 95% ethanol, phosphotungstic acid 0.015g/100ml) and 5 min in
EA50 (0.5% lightgreen SF yellowish CI 42095, 0.5% bismark brown CI 21000, 0.5%
eosin Y CI 45380, 0.2g phosphotungstic acid and 2 drops of lithium carbonate
(0.154g in 10ml dH2O)).