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. Author manuscript; available in PMC: 2021 Aug 19.
Published in final edited form as: J Am Chem Soc. 2020 Aug 4;142(33):14080–14089. doi: 10.1021/jacs.0c01064

Figure 6. Quadruple bioluminescence imaging.

Figure 6.

(a) Mutant luciferases were expressed in bacteria. Comparable levels of expression were observed (Figure S17). Cells were lysed, then distributed across four 96-well plates (in a 3×4 matrix). Saturating doses of the corresponding luciferin analogs were administered ([PhOH-Luc] = 250 μM, [7′-DMAMeLuc] = 250 μM, [4′-BrLuc] = 100 μM, [4′-MorphoLuc] = 250 μM). Images were acquired, and the raw emission data were stacked and unmixed. (b) Unmixed images from (a) depicting a quartet of unique luciferase-luciferin pairs. Emission intensities are plotted as total photon flux values. Error bars represent the standard error of the mean for n = 3 experiments.