Four samples of live LLC-Pk1 cells were treated with fluo-4 (a calcium sensitive fluorophore). Each sample was then exposed to a different stimulus while submerged in HBS buffer, Time lapse images were taken at 10X magnification for 120 s (1 frame every 2s) to capture the cellular response to the stimuli via fluorescent activity (Figure 3). Fluorescence intensity versus time data reveal patterns of fluorescence intensity (and therefore, cytosolic calcium level) fluctuation based on specific stimuli introduced to the extracellular space (Figure 1). Cells treated with HBS (control group) expectedly demonstrate no significant fluctuation in fluorescence intensity overtime, and therefore were not able to quantitatively analyzed any further (Figure 1), Cells treated with bradykinin appear to have a delayed rise in fluorescence intensity over time (44s, Table 1) whereas ATP treated cells appear to have an immediate and oscillating fluctuation in fluorescence intensity over time (3s, 18s, respectively, Table 1). Only a small number of cells responded to bradykinin introduction with an increase in fluorescence intensity, while a majority of the cells treated with ATP demonstrated a response (11/154, 97/99, respectively, Table 1). Additionally, the relative increase of fluorescence intensity in cells treated with bradykinin appears to be much larger than that of cells treated with ATP (589, 331, respectively, Table 1). Vasopressin treated cells demonstrated no fluorescent fluctuation over time, and therefore were not able to be quantitatively analyzed any further (Figure 1). Reasons for this absence of an expected response are detailed in the discussion.
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