Live Cell Multicolour Lifetime Imaging Using Genetically Encodable Fluorophores

Nine fluorescent proteins (FPs) simultaneous imaging has been demonstrated in a single acquisition using fluorescence lifetime imaging microscopy (FLIM) combined with pulsed interleaved excitation (PIE) for three laser lines. We also show how to unmix spectrally similar FPs in a pixel-by-pixel manner with an analytical non-fitting solution. ### Competing Interest Statement The authors have declared no competing interest.