Protocol for measuring fly sizes
Raise flies in uncrowded conditions
1. Let many flies lay eggs on agar plate+yeast paste for about 12h at 25C.
2. Let eggs develop for 24h at 25C. (Larvae resist better to the next step)
3. Collect larvae on a net and put about 30 larvae in a tube (with Kimwipe paper at the bottom)
4. Let the flies develop at 25C. Use only tubes from which less than 45 flies total hatch.
Wing size, palp size, leg size
1. Dissect 2-4-day old flies in 70% ethanol. One wing, one palp, one leg for each fly.
2. Put a drop of 25uL of Hoyer’s medium diluted in water (1:1) on a slide.
3. Put the 3 organs in the drop (using a 10uL pipet).
4. Put a cover slip.
5. Let it equilibrate for 1h.
6. Warm the slide for 24h so that the mounting medium polymerizes.
7. Use Alex’ script in IPlab to take the pictures+measurements.
Pupal case
1. store empty pupal cases in 70% ethanol.
2. align the pupal cases on a double-sided sticky tape on a slide, with ventral part facing the top.
3. remove the operculum.
4. take pictures with the stereomicroscope (no zoom). Save them as JPEG flies.
5. make measurements of pupal case area (without the operculum area)
and pupal case length using the script “pupal case - Vi” in IPlab.
Dry mass
1. Freeze 8-10-day-old flies in liquid nitrogen (5 flies per tube).
2. Dry the flies at 37C for 1 day.
3. Measure using
Glycogen, Lipid, InR Tyr kinase activity: 8-10-day-old flies