MMA Plastic embedding:

Week1:

(Mon-Wed) cut unneeded metaphysis off; 1-3 days 100% ETOH changes 4C – we do this in cassettes to save time with solution changes
(Thurs) Acetone 4C O/N
(Fri) Xylene over weekend 4C, in a desiccator

Week2:

(Mon) MMA1 stir 1hr, room temp. vacuum O/N; Make MMA2 stir 1hr, store 4C O/N (mix in the morning before use)
(Tue) MMA2 room temp. vacuum 8hr; O/N 4C; Make MMA3 stir 1hr, store 4C O/N (mix in the morning before use)
(Wed) MMA3 1-2hr vacuum on ice; O/N 4C; Make MMA3 stir 1hr, store 4C O/N (mix in the morning before use)
(Thurs) MMA3 2-4hr vacuum on ice; O/N 4C
(Fri) Make embedding MMA3 Fresh, stir for 1hr at 4C, embed, N gas 5min, -20 for weekend
- We superglue the bones in place to the bottom of the embedding molds to reduce movement.

Week3:

MMA solutions:

MMA1: 10mL/sample

MMA2: 10mL/sample

MMA3: 10mL/sample

MMA3 for embedding: 3mL/sample

Methyl methacrylate 60mL – 90mL
Butyl methacrylate 35mL – 52.5mL
Methyl benzoate 5mL – 7.5mL
PEG 400 1.2mL – 1.8mL
Benzoyl peroxide 0.8g – 1.2g
- Total about 100mL –150mL
Add 0.4mL (in 100mL) or 0.6mL (in 150mL) N,N dimethyl-p-toluidine just before embedding, enough time for a thorough mix is all that’s needed – 5 minutes or less

Catalog numbers:

Methyl methacrylate (M55909-1L, Sigma)
Butyl methacrylate (235865-1L, Sigma)
Methyl benzoate (M29908-500g, Sigma)
Polyethylene glycol 400 (PEG 400) (202398-500g, Sigma)
Benzoyl peroxide (179981-500g, Sigma)
N,N dimethyl-p-toluidine (41490-100mL, Sigma)

De-plasticizing:

3x 20min washes in 2-Methoxyethl Acetate (109886-1L, Sigma)