Everything’s routine now. The moment I open the door to the Bryan Research Building, a quick rush of AC floods over me. I click the “^” button, step in, click the “4” button, step out. I round the corner, smile and wave to Grace in front of me, plop down at my desk, turn right to say hi to my mentor Jenny. I check the board, scanning the pinned schedule that Jenny and I write on the past Fridays. It will be a mixture of different procedures: burr hole injection surgeries, perfusions, brain slicing, mounting brain slices, imaging, and coding, respectively. Everything must be done in order, and each step takes time, leading to weeks worth of waiting to obtain data.
Each procedure is intricate and cannot be rushed.
I usually begin my week with the first procedure: burr hole injection surgeries. I carefully drill a hole into an anesthetized mouse’s skull in order to inject fluorescent tags called tdTomato. After a mouse receives a viral injection in its brain, it takes at least two weeks for the virus to be expressed, or visible through a microscope.
The snowball effect begins.
After two weeks, the mouse is ready to be perfused, and the mouse’s brain must sit in PFA overnight. After a night, the mouse’s brain is rinsed with PBS three separate times in 15 minute intervals. After approximately an hour, the mouse’s brain must be submerged in 30% sucrose for at least a day. After a couple days, the brain is be manually sliced into delicate, thin slices and placed in PBS once more. After the brain is divided, the brain slices are meticulously mounted onto glass slides where they must properly dry overnight. After a night, the researcher must make time to image every brain slice under a microscope connected to a camera. After taking pictures of the injected brain slices, the data must be analyzed in old or new MatLab code.
After this long process to collect data from one mouse, something could have gone wrong at any stage. The injection may have been too deep or too far to the left, the brain may have been damaged during perfusion, the brain could have been sliced at the wrong angle, the injected area may have been physically stretched out during mounting, and more. Science is slow, and I never understood why researchers say this so often until now.
Despite the weeks it takes to collect data from one mouse, I’ve learned to appreciate all researchers who have brilliant ideas and work day and night to generate data that may or may not significant. I take Dr. Glickfeld’s words to heart: “We don’t ever hope to see a certain result.” Anything that comes out of experiments will contribute to science in some way.
I have a rhythm at work now where I can easily come into lab to get into a flow. I’m sad to think that BSURF only has three more weeks left, but I hope to hop back unto the Glickfeld Lab once the school year starts. I love the work here, and even though I don’t have a lot of neurobiology background, I am happy to learn something new everyday. I can’t wait to see what these next three weeks will bring!
