Manipulating mosquito eggs

Now that we have stable caged populations of Culex mosquitoes and have worked out an egg laying on demand system the next incremental step is to separate the individual eggs from freshly laid egg rafts for microinjection.  (We have been advised that injecting them while together in an egg raft does not work well because of the pressure they put on each other during injections...)  Jolene worked out the timing to get fresh rafts, disassociated them, and lined them up on wet blotting paper (to keep them from drying out) on a microscope slide.


To the immediate left of the eggs is an extra layer of paper to brace them for injection from the right.  They were very light in color at first but they got darker (and harden) quickly by the time the picture was taken.  She put this first set back into water to see if manipulating them affects survival.  (For more photos and news check out Jolene's research blog.)

In other related news, we have some of the plasmids we need cloned and I am working on cloning the last one this week (that contain the genetic modifications for injection), and my plasmid preps are working!  Fingers crossed, with any luck we can try serious microinjection to generate germ line Culex transformations three weeks from now during the next round of fresh egg production.  We are planning to generate the self docking system described (in Anopheles gambiae) by Meredith et al. 2013 in Culex.

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