A few steps back, but looking forward
As we move into the penultimate week of classes, I was looking forward to conducting my first morphological studies of Nrf3 knockdown fish exposed to MEHP.
Practicing microinjection has been an arduous journey, as a number of factors in technique can lead to the improper injection of a morpholino into a fish embryo. Ensuring that the size of the needle I’m poking embryos with and the pulse duration of the injector are enough so as to not damage the fish’s structures but also enough to properly incorporate into the fish’s genome is a tough balance to maintain. Just recently, however, I was able to achieve the proper efficiency necessary for conducting morpholino studies.
That is until, it was determined that our methods for MEHP exposure have not been standardized to our colleagues’ at the University of Massachusetts Amherst. It turns out, therefore, that a good deal of the work that I’ve completed this semester, and a portion of the work that was completed by Nancy, the thesis student before me, will have to be repeated.
While such a turn of events is frustrating at the outset, it is important for me to remain grounded in my work and consider the positives of this situation. First of all, I must recognize that these circumstances are simply the nature of this field that I’ve chosen to commit myself to. Experiments and techniques must be meticulously managed, and in many cases on this road, I will be forced to move into the breakdown lane. In this case, however, I’m going to have to actually get myself moving instead of calling AAA. (Did you know there was a five call per year limit on your AAA membership? Unfortunately I’ve already used all of mine for this year). The realism of this situation cannot be avoided, as in essence it is exactly what I signed up for; real experience working in an academic lab.
Additionally, a majority of the work that is derived from my predecessor on this project will require microinjection. Nancy’s work, like mine, was studying the effect of MEHP exposure on subjects who have had a specific transcription factor from the CNC-bZIP family knocked down by morpholino. Repeating her studies will simply ensure my further honing of my microinjection skills. Along with this, by picking up part of her research, I will actually be characterizing the entire Nfe2-related group of the CNC-bZIP family, and in this way my understanding of these transcription factors and the family as a whole will only improve.
While this turn of events definitely begets retread, I am not at square one. The techniques that I’ve picked up from this semester will carry into next semester and only improve. Additionally, I have an entire library of baselines for study, as I will have collected control groups for each of my studies by the end of next week. Immer weiter.