Whole Mount Preparation of Unstained Abdominal Cuticle of Adults

1. Preserve adult flies in 70% ethanol for 3-7 d.

2. Separate the abdomen in 70% ethanol and make either a mid-ventral or mid-dorsal longitudinal cut in it to obtain a complete dorsal or ventral view of the adult cuticular pattern, respectively.

3. Transfer individual pieces to separate Stender dishes with 70% ethanol. Using forceps and without damaging the cuticle, remove the internal organs and fat body as much as possible.

4. Pass the specimen through the descending grades of ethanol (70%, 50%, and 30%) and finally through distilled water, keeping the specimen in each of the solutions for 2 min (see Subheading 3.1., step 12).

5. Transfer the piece to a dish on a hot plate (50°C) containing 1 N NaOH and cook for 2-3 min. Now the tissues attached to the cuticle will become translucent.

6. Transfer the cooked integument to a dish containing distilled water. Under the dissecting microscope, holding the skin to the bottom of the dish, carefully squirt water several times to remove all the digested tissue remains.

7. Mount the clean cuticle in water mounting medium following Subheading 3.1., steps 23-27 (see Fig. 5A).

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