Abstract
Recent progress in molecular engineering of genetically encoded probes whose spectral properties are controlled with light, such as photoactivatable, photoswitchable and reversibly switchable fluorescent proteins, has brought the new possibilities to bioimaging and super-resolution microscopy. The development of modern photoconvertible proteins is linked to the studies of light-induced chromophore transformations. Here, we summarize the current view on the chromophore chemistry in the photocontrollable fluorescent proteins. We describe both the fundamental principles and the specific molecular mechanisms underlying the irreversible and reversible chromophore photoconversions. We discuss advancements in super-resolution microscopy that became possible due to the engineering of new protein phenotypes and understanding of their chromophore transformations.
🔬 Techniques
✨ Fluorophores
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📊 Figures
Figure 1
Light induced chromophore transformations in irreversibly photoswitchable FPs
Colors highlighting chromophores correspond to the spectral range of observed fluorescence. Non-fluorescent chromophores are shown in gray. [O] denotes an oxidation reaction. (a) Core chromophores of ...
Figure 2
Light induced chromophore transformations in reversibly photoswitchable FPs
Colors highlighting chromophores correspond to the spectral range of observed fluorescence. Non-fluorescent chromophores are shown in gray. (a-e) Chromophore transformations shown for Dronpa (a), Padr...
Figure 3
Proposed combinations of photocontrollable FPs for super-resolution microscopy
(a) Green PAGFP and red PATagRFP or PAmCherry1 are suitable for simultaneous two-color PALM. Both FPs are activated by violet light and fluorescence signals of activated FPs are detected in separate c...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
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