I must carry out a microinjection of dsRNA to the parasitic wasp Nasonia vitripennis, but sometimes I see that a large number of the injected individuals die after one or few days, even if I keep myself very careful during the task. Every week I use different breeding stocks, in some breeding stocks the mortality rate are very low after injection, whereas in other stocks show high mortality rate. I wonder why such variation occurs.
Some colleagues working on honeybee virus developed this tecnique for infection.
They anestethyzed the bee with CO2.
They shaved the scutum/scutellum completely using micro scissor and a microscope.
They brushed scutum/scutellum a little using a razor blade.
They dropped (a couple of microliter) of the solution with viralRNA and wait until dry.
I don't know if this can work for you.. but of course that can't be done on a large numbers of individuals.
I don't know which tool you use for dsRNA injection. I use Nanoject II from Drummond, which uses glass capillaries as needles, to inject dsRNA in mosquitoes. These capillaries are extented under a flame to obtain a very long and narrow needle. But then I have to break them to open the capillary... I noticed that the quality of the break is very important. If the break is not clean (for exemple beveled) I have a huge mortality. Same thing if the capillary is to wide... The wound on the insect is bigger and they die more.
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