Photoactivatable Fluorescent Dyes with Hydrophilic Caging Groups and Their Use in Multicolor Nanoscopy

We propose a series of fluorescent dyes with hydrophilic carbamate caging groups that undergo rapid photoactivation under UV (<= 400 nm) irradiation but do not undergo spurious two-photon activation with high-intensity (visible or infrared) light of about twice the wavelength. The caged fluorescent dyes and labels derived therefrom display high water solubility and convert upon photoactivation into validated super-resolution and live-cell-compatible fluorophores. In combination with popular fluorescent markers, multiple (up to six)-color images can be obtained with stimulated emission depletion nanoscopy. Moreover, individual fluorophores can be localized with precision <3 nm (standard deviation) using MINSTED and MINFLUX techniques.

Automated summary

The study introduces a series of fluorescent dyes with hydrophilic carbamate caging groups that can rapidly photoactivate under UV irradiation and are not affected by high-intensity light of double their wavelength. These dyes have high water solubility and transform into validated super-resolution and live-cell-compatible fluorophores upon photoactivation. Using popular fluorescent markers, multiple color images can be obtained with stimulated emission depletion nanoscopy, while precise localization of individual fluorophores can be achieved with a standard deviation of <3 nm using MINSTED and MINFLUX techniques.