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17 March 2023 Photoactivatable fluorophores for bioimaging applications
Yang Zhang, Yeting Zheng, Andrea Tomassini, Colin E. Hayter, Françisco M. Raymo
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Proceedings Volume 12395, Colloidal Nanoparticles for Biomedical Applications XVIII; 1239503 (2023) https://doi.org/10.1117/12.2647135
Event: SPIE BiOS, 2023, San Francisco, California, United States
Abstract
Single-molecule localization microscopy (SMLM) strategies based on fluorescence photoactivation permit the imaging of live cells with subdiffraction resolution and the high-throughput tracking of individual biomolecules in their interior. They rely predominantly on genetically-encoded fluorescent proteins to label live cells selectively and allow the sequential single-molecule localization of sparse populations of photoactivated fluorophores. Synthetic counterparts to these photoresponsive proteins are limited to a few remarkable examples at the present stage, mostly because of the daunting challenges in engineering biocompatible molecular constructs with appropriate photochemical and photophysical properties for live-cell SMLM. Our laboratory developed a new family of synthetic photoactivatable fluorophores specifically designed for these imaging applications. They combine a borondipyrromethene (BODIPY) fluorophore and an ortho-nitrobenzyl (ONB) photocage in a single molecular skeleton. The photoinduced ONB cleavage extends electronic delocalization to shift bathochromically the BODIPY absorption and emission bands. As a result, these photochemical transformations can be exploited to switch fluorescence on in a spectral region compatible with bioimaging applications and allow the localization of the photochemical product at the single-molecule level. Furthermore, our compounds can be delivered and operated in the interior of live cells to enable the visualization of organelles with nanometer resolution and the intracellular tracking of single photoactivated molecules.
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Yang Zhang, Yeting Zheng, Andrea Tomassini, Colin E. Hayter, and Françisco M. Raymo "Photoactivatable fluorophores for bioimaging applications", Proc. SPIE 12395, Colloidal Nanoparticles for Biomedical Applications XVIII, 1239503 (17 March 2023); https://doi.org/10.1117/12.2647135
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KEYWORDS
Fluorophores
Fluorescence
Live cell imaging
Optical tracking
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