New GFP Variants with Enhanced Two-Photon Brightness


Rosana S. Molina, Robert E. Campbell, Nathan C. Shaner, Thomas E. Hughes, Mikhail Drobizhev


Biophysical Journal


The two-photon (2P) brightness of fluorescent proteins (FPs) is crucial for 2P microscopy. Bright proteins mean that more images can be collected with less heating and damage. We know that the optical properties of a specific chromophore are shaped by the local electrostatic environment created by the surrounding protein. Here we show that as the one photon absorption (1PA) peak position in a series of FPs - with the same anionic GFP chromophore - shifts to the blue, the peak 2P cross section and 2P brightness increases.



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