Luminescence exerts an ideal optical readout for imaging living subjects including no external light source, whereas the dim luminescence and poor color pallet should be addressed for the better utilities. To address the demerits and to prevail the advantages, we developed a bright luminescent protein, named yellow Nano-lantern, exhibiting about 10-20 times brighter than wild-type RLuc. In this chapter, we demonstrate two luminescence-based protocols in detail: i.e., (a) multicolor visualization of Ca(2+) dynamics in different cellular compartments in a single cell using Ca(2+) indicators based on cyan- and orange-Nano-lanterns and (b) video-rate tumor detection in a freely moving mouse using yellow Nano-lantern.
Keywords: Bioluminescence; Bioluminescence resonance energy transfer (BRET); Calcium dynamics; Renilla luciferase; Tumor imaging.