Skeleton preparation

From: Richard Behringer

Day 1.

• Take off the skin, remove viscera.
• Fix the mice overnight in 95% ethanol.

Day 2.

• Stain in alcian blue (0.05% alcian blue GX, 5% acetic acid) overnight.
• Put back in ethanol 2–5 hours.
• Treat with 2% KOH for 24h (until mostly clear) in a petri dish. Adults can be treated for 2 days or longer. At this point, remaining fat and skin can be taken off more easily.

Day 3.

• Stain overnight in 1% KOH, 0.015% alizarin red. (Be careful while moving the mice - use slide).

Day 4.

• Skeletons are cleared in 1% KOH, 20% glycerol for 2 days or more.

Day 5-7.

• Store skeletons in a 1:1 mixture of 95% ethanol and glycerol.