My Disappearing Vision and the Wrong Kind of Wasp!!

Wow! Intensive microscope stuff yesterday and today have got me slowly losing my ability to see in the dark. Anyway, it all gets healed over the weekend, and I haven't had a headache since the first week, so I must be adjusting.

So, yesterday, I worked on one large, important assignment: leaf disk density modification. I'll start from the beginning. We are using these leaf disks as breeding grounds for new wasps. The females that were mated on Monday were put onto these leaf disks today, but I'll get to that later. Anyway, leaf disks are made by cutting circular portions of leaves and are stored on small, agar filled Petri dishes. Those leaf disks initially contain about 100 (but in one case, 189) whitefly larvae of all instars (first through fourth). The density needs to be lowered to 50 second instar larvae. Of course, the tedious part is counting. The count is done in two parts. First, the larvae too close to the edge of the disc, and the ones that are obviously not second instar larvae (the first and fourth) are removed, along with any thirds, although they're sometimes harder to differentiate. After that stage, the leaf is mapped, and a count of all of the whiteflies is done by section of the leaf, which greatly simplifies the process. After that number is determined, whiteflies are removed until only 50 are left. This whole process takes me up to an hour per disk, but Suzanne is able to do three times that!

Today, we worked on creating the final disks (we needed fifteen in total), and we placed the mated females onto the disks. That part is kind of difficult because it requires the ability to see a tiny speck of a wasp drop down into a dish, and ensure that it doesn't escape (although fortunately, the females tend to want to stay on the disks and oviposit). The worst part is that there's no way to really check whether or not the wasp has landed past an initial, 5-second inspection, because the dish needs to be immediately covered up. For the rest of the day, we worked on isolating Eretmocerus emiratus pupae for my experience. Unfortunately, what I thought were Rickettsia-negative, Wolbachia-positive wasps were actually Rickettsia-positive, Wolbachia-negative wasps. That's bad, in case you were wondering. Fortunately, at another lab bench, the correct wasps were being harvested, and since we only need the males, and it doesn't matter whether or not they've mated when we use them, a gigantic crisis was averted. All we need to do is aspirate some males and that'll be all. We do still have to do a DNA extraction on all of the E. emiratus we used today, in order to be sure of what happened, since infection status can't be seen with the naked eye.