Visualization of subcellular organelles in vivo is critical for basic biomedical research and clinical applications. Two new flavonoids with an amide substituent were synthesized and characterized. The flavonoids were nearly non-fluorescent in aqueous environment, but exhibited two emission peaks (one λem at 495-536 nm and the other at 570-587 nm) in organic solvents, which were assigned to the excited normal (N*) and tautomer (T*) emission. When the dyes were examined on oligodendrocyte cells, they were found to selectively accumulate in the endoplasmic reticulum (ER), a eukaryotic organelle involved in lipid and protein synthesis, giving fluorescence turn-on. The ER-selective flavonoids could be a valuable tool due to its low molecular mass (<500), large Stokes' shift, low toxicity, and biocompatibility.