Alcian green stain for cartilage
This is an easy method for stianing cartilage that can be used with isolated or CAM-grafted limbs to study limb development or with whole embryos (8 to 12 days of incubation, ).
1. Fix limbs or embryos in 5% TCA for 1 hour at room temperature.
(Eviscerate embryos before fixation).
2. Wash 3X with PBS.
3. Immerse in Alcian green stain for 4 hours with gentle agitation.
4. Wash 3X with 70% ethanol, several hours to overnight.
5. Wash with 85%ethanol, 95% ethanol, 100% ethanol, several hours to overnight each.
6. Clear by washing 1-3X with methyl salicylate.
Alcian green stain:
0.1 g Alcian green
70 ml 100% ethanol
1 ml conc. HCl
29 ml dH2O
(100 ml)
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