Today marks the end of the first week of my Laidlaw research project. There was a lot that was accomplished and learned, one of the things being the importance of the fine motor control of the hands when dissecting specimens.  

My first three days, namely Monday, Tuesday, and Wednesday, were spent practicing dissection on Drosophila melanogaster flies. The days followed the same old, dull routine (yes, I am referring to Escape (The Piña Colada Song) - you will see why): coming into the lab in the morning, going into the fly room, putting on a lab coat, setting up slides, choosing pupae from fly tubes, fixing them on the tape with tweezers, transporting the slides to a separate lab room, and proceeding to dissect them using a scalpel. Thus, memorising song lyrics that I occasionally could hear from the radio in one of the lab rooms was a way to boost morale when I ended up dissecting 30 flies I spent an hour dissecting only for none of them to be suitable for staining.

Figure 1: D. melanogaster pupae on a glass slide.

Moreover, when I had the occasion to go outside to sift through dirt obtained from potted plants in the write-up area I was thrilled to be outside for a bit, although the weather on those days was not very summer-y. 

Figure 2: In search of suspected Bradysia sp. fungus gnat pupae, larvae, and/or adults in the soil collected from plant pots.

Then came Thursday, when my supervisor and I travelled to the University of Edinburgh to collect three batches of Anopheles sp. mosquitoes which were then transported in ice cream containers on a public train (a very exciting experience indeed). Once we got them safely into the lab, the real work began. Thursday afternoon, Friday, Saturday, and Sunday were all spent on adapting to cutting the soft bodies of the larvae and the pupae that did not have a pupal case (unlike the D. melanogaster pupae which I had practiced on). Once I cut the larvae and pupae, I stained them and then proceeded with the visualization of the cut specimens using a confocal microscope. Unfortunately, none of the images can be used in the final essay; however, the photographs of the specimens helped me understand how to improve my dissection and learn the morphology of the specimens (and, they were so pretty to look at!). 

Figure 2: Anopheles sp. mosquito pupa that has shed its larval molt. The photo was taken using a light microscope on an iPhone camera.

Overall, from running around the office with a fly tube in order to catch adult fungus gnats to learning how to dissect pupae the size of a splinter to transporting mosquitoes on a public train, in such a short amount of time I have accumulated many anecdotes, with hopefully many more to come.

On a more serious note, I already feel more confident as a researcher because I was able to practice keeping a lab notebook, calculate the dilution factor for the staining dye (Hoechst 34580), and learn how to wash, fix, stain, and visualise dissected pupae. Furthermore, I attended a group meeting where I learned what research is currently being carried out by a PhD student on the membrane microdomains present in D. melanogaster. During that meeting I contributed by discussing the progress I was making in culturing a new population of fungus gnats (which might potentially also be used in my research). The lesson learned was that it is possible to grow fungus gnats in an erlenmeyer with soil and a bit of honey. 

As I embarked on this new journey of learning about abdominal metamorphosis in insects other than D. melanogaster, something that is quite under-researched, I felt both excitement and anxiety because in order to get results I must master dissection in under 6 weeks, a skill that often takes months of practice. Moreover, though there are a few published research articles on metamorphosis in mosquitoes, there are not many sources which I could use to figure out the timing of development and the pre-pupation IV instar in Anopheles in particular. Therefore, I must somehow observe and capture the morphology of as many larva as possible next week to understand which larva are the most likely to pupate, with pupation serving as time 0. 

What I have accomplished this week was gaining enough experience to dissect the pupae and larvae specimens I will use in the upcoming weeks for my research in addition to practicing fixation with paraformaldehyde, washing the specimens with PBS buffer, understanding the conditions needed for the Hoechst staining dye to properly label the nuclei as well as the effects of different solvents on tissue, and becoming more familiar with identifying morphological structures. 

My goals for next week are to find several larvae directly before they turn into a pupae, continue working on improving my dissection so as to preserve as much of the tissue as possible, and start working on understanding the timing of development to be able to produce time-accurate results.