Emissary Akash Visits the Kuo Lab at Loyola University Chicago

I spent a week in November 2018 in Dr. Ivana Kuo’s laboratory (Loyola University Chicago) as an Emissary of the NSF NeuroNex Bioluminescence Technology Hub. Dr. Kuo’s lab routinely uses fluorescence Ca2+ imaging, but is considering applications for which bioluminescent Ca2+ imaging would be advantageous. Since the lab had not done bioluminescence imaging before, we started by imaging light emission from HEK cells stably expressing a Gaussia luciferase construct. We routinely use this cell line to “baseline” the imaging set-up (robust signal, dependent on coelenterazine (CTZ) concentration, absence of any contaminating outside light). We then proceeded to image cells transiently expressing a Ca2+ dependent split luciferase.

We used HEK293 cells and C2C12 muscle cells, stimulated calcium flux with electrical pacing, Isoproterenol or Ionomycin, varied CTZ concentrations and recording buffer compositions, as well as exposure times and intervals. It was an intensive 4 days of experiments that allowed both myself and the members of the Kuo lab to focus on systematically testing experimental parameters. I went back to my home lab with a list of experiments to carry out under my experimental conditions for comparison to the results we got in the Kuo lab.

While at Loyola University Chicago, I also had the chance to give a talk on my Ph.D. work. Overall, my visit to the Kuo lab was a fantastic opportunity that took me outside my comfort zone and helped me grow as a graduate student.

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