Lab #11 – CaMPARI Validation Time Course

In this lab, we investigated the fluorescence response of CaMPARI-expressing cells to two treatments: ATP and Tris-HCl, which will serve as controls for our pharmacology experiment next week. ATP is expected to increase intracellular calcium levels, which should cause a fluorescence shift in CaMPARI due to its calcium-dependent photoconversion. Tris-HCl serves as a negative control and is not expected to alter intracellular calcium levels, so no significant change in fluorescence is anticipated. Observing fluorescence over time, rather than at a single point, is crucial to determine how quickly or slowly the calcium response occurs, which informs us about the dynamics of CaMPARI activation and the biological relevance of each treatment in a time-dependent pharmacological context.