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bioRxiv · 10.1101/2021.05.20.444797

Systematic Tuning of Rhodamine Spirocyclization for Super-Resolution Microscopy

Lardon, N.·Wang, L.·Tschanz, A.·Hoess, P.·Tran, M.·D'Este, E.·Ries, J.·Johnsson, K.
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Abstract

Rhodamines are the most important class of fluorophores for applications in live-cell fluorescence microscopy. This is mainly because rhodamines exist in a dynamic equilibrium between a fluorescent zwitterion and a non-fluorescent but cell-permeable spirocyclic form. Different imaging applications require different positions of this dynamic equilibrium, which poses a challenge for the design of suitable probes. We describe here how the conversion of the ortho-carboxy moiety of a given rhodamine into substituted acyl benzenesulfonamides and alkylamides permits the systematic tuning of the equilibrium of spirocyclization with unprecedented accuracy and over a large range. This allows to transform the same rhodamine into either a highly fluorogenic and cell-permeable probe for live-cell stimulated emission depletion (STED) microscopy, or into a spontaneously blinking dye for single molecule localization microscopy (SMLM). We used this approach to generate differently colored probes optimized for different labeling systems and imaging applications.

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Lardon, N., Wang, L., Tschanz, A., Hoess, P., Tran, M., D'Este, E., Ries, J., Johnsson, K.. 2021-05-20. Systematic Tuning of Rhodamine Spirocyclization for Super-Resolution Microscopy. https://doi.org/10.1101/2021.05.20.444797

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