While fluorescent proteins are powerful tools, they have some limitations: - Photobleaching: Fluorescence can diminish over time when exposed to intense light. - Phototoxicity: Prolonged exposure to light can damage living cells. - Size: The relatively large size of fluorescent proteins can potentially interfere with the function of the fusion protein. - Spectral Overlap: Emission spectra of different fluorescent proteins can overlap, complicating multicolor imaging.
