Visible Light Bioluminescence Imaging Platform for Animal Cell Imaging.

The present protocol introduces a visible light bioluminescence imaging (BLI) platform together with 12 novel coelenterazine (CTZ) analogues and luciferase sets. We exemplify to create diverse hues of bioluminescence (BL) ranging from blue to far red with the combination of marine luciferases and the three groups of CTZ analogues. We also show how to characterize the new CTZ analogues in detail such as the kinetic parameters, dose dependency, and luciferase specificity. The 2-series CTZ analogues interestingly have specificity to artificial luciferases and are completely dark with Renilla luciferase derivatives in contrast. The 3d is highly specific to only NanoLuc. This BL imaging system covering the visible region provides a useful multicolor imaging portfolio that efficiently images molecular events in mammalian cells.