I need to micro-inject (nanoject, 13-32 nl) sand fly larvae with a solution and it is very difficult to assess whether or not it has been injected, so I need to mix the solution with some sort of colorant. Evans blue, perhaps?
Evans blue is a god choice. My lab has used it extensively for in vitro work, and we have not encountered any toxic effects. We used it at 1%. It may bind to hemolymph proteins, and thus could possibly enter cells by receptor mediated endocytosis. We have never encountered this, even though we had active oocytes in these studies. There are several references, but the most important is
. Davey KG & Huebner E (1974) The response of the follicle cells of Rhodnius prolixus to juvenile hormone and antigonadotropin in vitro. Can J Zool 52: 1407-1412.
You are wise to seek reassurance that such small volumes have actually entered the insect. On some occasions I have partially filled the needle ( finely drawn glass pipet) with oil and then I draw up the precise volume to be injected. I then inject slowly to the oil-solution interface. Of course this only works for materials that are insoluble in oil.
I am VERY impressed that you are injecting such tiny organisms!
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