Firefly shows bioluminescence by “luciferin−luciferase” (L−L) reaction using luciferin,
luciferase, ATP and O2. The chemical photon generation by an enzymatic reaction is widely utilized
for analytical methods including biological imaging in the life science fields. To expand photondetecting
analyses with firefly bioluminescence, it is important for users to understand the chemical
basis of the L−L reaction. In particular, the emission color variation of the L−L reaction is one of the
distinguishing characteristics for multicolor luciferase assay and in vivo imaging. From the viewpoint
of fundamental chemistry, this review explains the recent progress in the studies on the molecular
mechanism of emission color variation after showing the outline of the reaction mechanism of the whole L−L reaction. On
the basis of the mechanism, the progresses in organic synthesis of luciferin analogs modulating their emission colors are
also presented to support further developments of red/near infrared in vivo biological imaging utility of firefly bioluminescence.
Keywords: Firefly, Bioluminescence, Luciferase, Luciferin, Multicolor assay, NIR spectroscopy, In vivo imaging.
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