# Muscle mass measurement
[EcoEvoDevo Lab](https://hackmd.io/@EcoEvoDevoLab/AngeliniLab)

For measurement of flight muscle mass in true bugs. Based on the protocol of [O’Brien & Boisseau 2018](https://doi.org/10.1371/journal.pone.0206997).
::: info
It may also be possible to use this protocol to determine the dry mass of the gonads.
:::
- [ ] Image newly molted adult bugs. Be sure to apply a unique ID number.
- [ ] Euthanize and store at -20˚C
- [ ] Using a sharp scalpel, remove the head and legs (at the coxa). Dissect away the abdomen. -- Be careful to cut such that the dorsal and ventral boundaries of the thorax and abdomen are separated.
- [ ] Dry at 70˚C (overnight at least)
- [ ] Weigh the sample using an accurate balance (in the Johnson lab area)
- [ ] Fully submerging the tissue in 10% KOH
- [ ] Incubate at 70˚C for 1-8 h
::: warning
My guess is that 1 h is too short, and 8 h is too long. Try 4 hours!
:::
- [ ] Dry at 70˚C (overnight at least
- [ ] Weigh a second time
The difference between the first and second weighing is an estimate of dry muscle mass.
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[EcoEvoDevo Lab](https://hackmd.io/@EcoEvoDevoLab/AngeliniLab)