Trehalose measurement in Würzburg with Dennis Pauls

Draft of protocol

Material:

0.5 µl capillary with adequate pump, 0.5ml and 1.5ml Eppis, ice, peaked stylus, razor blade, 2 forceps (or scissors)

Procedure:

sting 3 holes in a small eppi (size is important, not to small so that enough hemolymph can go through, not to big so that no fat or other dirt is going through), put into big eppi.

20 adult flies on ice, cut their wings (not sure about the reason)

spear the fly’s thorax with the peaked stylus

store flies in the small eppi with holes

centrifuge the small eppi within the big one (1 min, 5000 rpm)

throw away small eppi, cut big eppi (to access the pellet easier), soak the pellet with a capillary

record the amount of soaked hemolymph with a ruler (e.g. by photographing under disscetion scope)

pump the hemolymph out of the capillary into a fresh small eppi and freeze it.

Remarks:

Everything has to be done on ice, cooling chain should as poosible not be interrupted

(5000 rpm can be differ with different centrifuges…)