14.4.2. Conditioning the bees

1. Place the conditioned stimulus (CS; disc with vertical pattern) into the central holder of the box.

2. Place a drop of the sucrose reward (e.g. 50 % sucrose) in the opening of the disk (Fig. 38 C).

3. Let the marked bee visit the CS pattern with the reward for five times, before starting with the first dual-choice test.

4. After five conditioning trials, test the choice behaviour of the bee by

4.1. Cover the centre with a neutral white disc (Fig. 38C)
4.2. Present the CS pattern and an alternative test stimulus (e.g. a horizontal pattern) to the left and to the right side of the box.

5. Register all approaches towards the left and the right stimulus for 1 min.

6. Then reverse the side of the tested stimuli and record the choice behaviour of the bee again for 1 min.

This procedure helps to avoid artefacts due to a side preference of the bee.

7. Afterwards, condition the bee again five times to the CS pattern.

8. Repeat the dual choice test to both the vertical and horizontal stimuli.

9. Exchange each disc with the stimulus after one presentation to avoid scent marks of the bee; discs or substrates used for conditioning should not be used again for the testing.