When you read a research article, what you see is a finished product. Behind that finished product are countless hours, weeks, months, and likely years of time and effort that went into making that published article. Not only do the experiments take time, but so does setting up and troubleshooting all of the techniques that were used. In science, things rarely work the first time you try them; machines will break, programs will crash, your entire procedure could fail. This is why one of the most important, and often undervalued, skills in science is being able to troubleshoot when things go wrong.
One of the primary components of my research is Gad1, which is a gene that codes for GABA, the primary inhibitory neurotransmitter in the brain. In order to target this gene, I had to be able to stain the brain tissue I was using to visualize Gad1. I quickly figured out this was not an easy feat.
Each of these pictures shows neurons from different rat brains. In each of these images, the blue is a marker for all neurons. The purple and red in the first three images show attempts to stain for Gad1 using immunohistochemistry (IHC). IHC is a stain for proteins, which works well to stain for other cell types such as dopamine and serotonin. Unfortunately, IHC does not work well for GABA. As you can see in these images, each of these stains has high background staining, with few actual neurons stained. After troubleshooting this technique for months, I decided to switch approaches and use Fluorescent In Situ Hybridization (FISH), which targets mRNA instead of proteins, instead. The final image shows successful Gad1 staining in pink. Here you can clearly see the GABAergic neurons with almost no background staining. This successful Gad1 staining took nearly a year of work between troubleshooting the IHC, switching to FISH, and tweaking the FISH protocol. As a scientist, one of my jobs is to find the most effective ways to conduct experiments, and in order to execute effective science you need to be able to troubleshoot when things go wrong. So, the next time you see a research article, keep in mind that you are seeing the final product. As in life, things in science don’t always go as planned. When this happens, shake it off and just keep swimming.
Contributed by Maddie Ray, a third year PhD graduate student at Boston College, and our Featured Artist for the fall of 2018. To meet Maddie and see more of her art, click here.
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hello zebrafish are cool