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Clearing with BABB

BABB provides greater clearing power than glycerol, and so is useful for clearing large or opaque tissues. However, the reaction products of alkaline phosphatase dissolve in BABB, so tissues should be photographed, and then returned to aqueous media by reversing the procedure.

1. Dehydrate embryos through a MeOH/H2O or EtOH/H2O series to 100% alcohol.
2. Transfer to 1:2 BABB to alcohol for approximately 20 min.
3. Then to 2:1 BABB to alcohol for approximately 20 min.
4. Into BABB for approximately 20 min.

Notes

• BABB is 1:2 benzyl alcohol to benzyl benzoate.
• Warning: embryos are very brittle in BABB.
• Warning: BABB is a nasty organic, so wear gloves.
• Must be done in polypropylene or glass. View in glass dishes only.

A Conlon Lab Protocol

pdf file for this protocol