2.1 Sample Collection

1. Anesthetize bees by keeping them at 4 ºC for a few minutes.

2. Mount them on a wax plate by a pair of insect pins crossing over the waist.

3. Incise dorsally between the 5th and 6th abdominal segment by means of a G 30 needle (BD).

4. Collect 1 µl of haemolymph using a microcapillary.

Care needs to be taken to avoid contamination from intestine and other surrounding organs.

5. Transfer each haemolymph sample into a 1.5 ml Eppendorf tube.

6. Immediately snap freeze the samples in liquid nitrogen.

7. Store them at -80 ºC until testing.