18.104.22.168. Dissection of the RCC from the head capsule
1. Sedate a live adult bee by chilling.
2. Decapitate the bee by gently pulling on the head to elongate the neck.
3. Sever the neck as close to the thorax as possible.
4. Secure the head, face side down, to a wax surface using a pair of pins through the eyes.
Angle the pins away from the head to maximize accessibility (Fig. 45.1).
5. Using a micro-scalpel or –scissors, cut the area (postgena) that encircles the occipital foramen.
6. Widen the circle out to the inner margins of the eyes.
7. Gently tilt up the cuticle from the anterior end to gain access to any connective tissue bound to the cuticle.
8. Sever these connections.
Be sure to minimally disturb the posterior end and leave the oesophagus intact.
9. Remove the cuticle plate.
10. Add a droplet of incubation medium if dissecting to measure the rate of juvenile hormone biosynthesis by the paired corpora allata (see section 3.5 of the BEEBOOK paper on physiology and biochemistry (Hartfelder et al., 2013)) or another appropriate physiological liquid (e.g. Ringer solution, see Table 1 of the BEEBOOK paper on cell cultures (Genersch et al., 2013)) to ensure the interior of the head stays moist (Fig. 45.2.).
11. Examine the posterior end of the brain where the suboesophogeal ganglion completes a circle around the aorta and oesophagus; the retrocerebral complex will be found in this area, also tightly associated with AO-ES.
12. If the glands are not immediately visible, gently pull on the oesophagus and the complex should come into view.
The RCC has a slightly blue colour that is distinct from the whiter nervous tissue around it (Fig. 45.3.).