Bleaching and clearing embryos
• Fix with Bouin's Fix at room temperature overnight.
• Wash with 70%EtOH-TE three times for 10 min.
• Incubate the embryos in bleaching solution (1%H 2 O 2 , 5% formamide, 0.5xSSC) under light at room temperature with rocking for 1 hour or until adequate bleaching is obtained.
• Dehydrate the embryos by incubating in two changes of methanol (5 minutes each).
• Transfer the embryos into glass slides or chambers and remove the methanol and replace it with 250
m
l of 2:1 BB/BA, and once fresh BB/BA.
BB: benzyl benzoate
BA: benzyl alcohl
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