Brainbrow Imaging of the Zebrafish Lateral Line: Retrospective

As the summer winds down, I have been reflecting on the project Dr. Steiner and I began working on in June. Dr. Steiner and I have been utilizing multi-transgenic zebrafish to deeply explore the regeneratory process of zebrafish sensory hair cells. The current body of zebrafish research suggests supporting mantle cells divide to produce hair cells during regeneration of neuromast sensory organs along the lateral line, although evidence of such is lacking. Together, we created a plan to manipulate three transgenic lines to image the regeneration of these neuromast cells, with the end-goal of better understanding this regeneratory process and the factors that control it.

With these research goals in mind, I’ve been learning about the complexities of conducting biological research with living specimens. Many times during the semester, my colleagues and I have sacrificed time to come in on weekends to feed the fish; there have been several weeks the fish didn’t cooperate with breeding, and we had to quickly work on a new plan for the week. Truthfully, though, working with live zebrafish brings an element of physicality and life to my research. It’s a pleasure starting my day seeing our entire population of zebrafish greet me with frantic swimming.

Also challenging, but extremely rewarding, is developing the Zebrabow process I’ve been using to visualize cells of zebrafish. At its best, this method makes the cells of zebrafish fluoresce a beautiful mosaic of colors, allowing a researcher to better understand the cell divisions that potentially lead to new hair cells. No single procedure will work for every lab; Dr. Steiner has greatly helped me in creating a Zebrabow process that works for our fish in particular. At the start of the summer, we had very little mosaic fluorescence amongst the cells. This provided me several opportunities to review the process we had used, and modify it for better performance in the future. Such modifications make this research project feel like it is truly our own; over the next few semesters, I hope to refine this procedure to obtain consistent and repeatable results.

This summer project has not only taught me about valuable lab techniques, such as confocal & fluorescent microscopy and taking care of living specimens; working with Dr. Steiner this summer made me increasingly comfortable with the research process and environment, which I hope to be a part of for years to come. Being able to pave my own way through a new set of procedures is fantastic experience for my future endeavors. Most of all, I’m excited to continue my research with Dr. Steiner over the upcoming year, building upon everything I’ve learned over the past months.

Included in this blog post is an image of one of the Zebrabow treatments I’ve done, in order for readers to better understand the process. This image was taken as numerous ‘slices’ of images from the confocal microscope, then compressed into a single image. On the bottom right is the neuromast, the organ containing hair cells that Dr. Steiner & I study. One can see numerous flourescent cells, as well as some macrophages (depicted by amorphous green-flourescent projections).