The second week at the lab definitely blew by. I think I’ve pretty much settled in and the days pass by so much quicker when things become routine—I’m no longer exerting as much conscious effort when it comes to typical lab work. The optogenetics protocol I mentioned in my last blog post is still being worked out, so for most of the week, I’ve been working on mounting rat brains. A little bit about mounting brain slices, after we run behavioral experiments on our rats and screen for different phenotypes (observable characteristics that result for gene x environment interaction), we extract their brain and process them.
The raw brain is very fragile and thus highly sensitive to most forms of mechanical manipulation. With that in mind, to give it more structural integrity as well as protect it from biological degradation, we perfuse it with formaldehyde (used in the embalming process). From there, we immerse the newly fixated brain in increasing gradients of sucrose solution (essentially antifreeze) over the course of several days so that we can keep the brain in subzero temperatures without it freezing.
Eventually, we run it through another series of steps (that I will describe in another blog post) that ultimately results in coronal cross-sections of almost the entire rat brain—each slice can be thinner than a human hair! These are then mounted via paintbrush (because of their thinness and fragility) onto microscope slides. This is often one of the most technically challenging aspects of lab work and it’s a standard Flagel Lab rite of passage. You receive the brain slices out of order since there’s no efficient way to keep the brain sections in the correct sequence during the cutting process. Thus, mounting the brains on the slides can prove to be a formidable task both mechanically and intellectually. Essentially, it’s like one complicated biological jigsaw puzzle. Moreover, when you’re still getting the hang of things, mounting can definitely be infuriating at times. The brain slices may run off the slide or you can accidentally sort one slice out of order and then have to take off all the following slices off the slide and remount them.
Yet, whenever I mount brains, I’m always reminded of those movies where some arcane martial arts master (who for some reason, only speaks in parables) assigns seemingly menial tasks to their student only for the student to later realize that those tasks served as the foundation for essential skills in martial arts. Similarly, brain mounting teaches you a lot regarding patience and the nature of research in general. As cliche as it may sound, patience is a virtue and persistence is key. There’s a lot of unglamorous work in research—for every graph or even point on a line someone had to put in hours of work running the experiment, processing the data, and potentially rerunning procedures just to fine tune the results. For the most part, research is more of a marathon than a sprint.
On a similar note, although the modern world moves at a blindingly fast speed, time works differently in the research setting. It serves as its own microcosm within the larger forces at work in the universe. There is still progress, but just imagine all of human knowledge as a circle that’s infinitely large, with each researcher being one infinitesimally small point on the circle pushing out, slowly but surely expanding the circle. As we advance, the absolute amount of progress each person can contribute remains fairly stable, but when we zoom out, we make less and less progress relatively as the circle expands—the law of diminishing returns. Occasionally we’ll get a breakthrough; one point on the circle pushes out, creates a sort of ripple effect, pulling out on nearby points of the circle. This tension on the rest of the circle allows the rest of the field to catch up and quickly reshape into a larger circle. The beauty of research or more generally, science, is that it is simply a series of trial and error processes built on top of each other. It brings out the kidlike curiosity we all once had, instead of pushing the boundaries of our understanding in our parents’ kitchens, we can now contribute to the collective knowledge of humankind in upgraded kitchens that we call laboratories. Connecting this back to my main point, although mounting brains might not be the most exciting thing in the world, progress is still progress. These brains I’ve mounted will undergo further chemical manipulation to screen for molecules of interest (proteins, receptors, and mRNA) in certain regions of the brain (PrL, PVT, NAc) and ultimately contribute to our knowledge regarding the neurobiological basis of addiction. Knowing that is enough for me to keep trudging onwards. Excelsior.
Until next time,