Wavelength-Sequential Generation of Multi-Emissive Fluorophores for Site-Selective Fluorescence Nanoscopy
Abstract
Photoactivatable fluorophores are indispensable tools for super-resolution microscopy, but their performance is often hindered by the need for caging group chemistry, ultraviolet activation, interfering triplet-state photochemistry, and most importantly resulting in a single emissive scaffold. Here, we report a hydroacridine-based fluorophore that addresses the often encountered limitations by providing caging-group-free activation using visible light. A heteroatom-engineering strategy enables rapid singlet-state proton-coupled electron transfer, converting a blue-emissive precursor into two highly fluorescent acridinium states with distinct green (ΦF ≈ 0.80) and red (ΦF ≈ 0.78) emission by a two-step photoconversion using blue and green light sequentially. The process operates efficiently under physiological conditions without involving triplet biradical intermediates, enhancing compatibility with live-cell imaging. The fluorophore architecture further supports organelle-targeted imaging and photoinduced intracellular redistribution, enabling multicolour visualization from a single molecular scaffold, serving both spatial and temporal control over the photoactivation. These findings provide new strategies for heteroatom-controlled visible-light photoactivation and fluorophore design, providing a new platform for next-generation super-resolution imaging.
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BibTeX
@misc{mondal2026wavelengthsequential,
title = {{Wavelength-Sequential Generation of Multi-Emissive Fluorophores for Site-Selective Fluorescence Nanoscopy}},
author = {Saugat Mondal and Tapas Bera and Debjit Maity and Sayan Paul and Swapnamoy Sahoo and Bhaskar Rao Pinninti and Mahitosh Mandal and Hannes A. Houck and N. D. Pradeep Singh},
howpublished = {ChemRxiv},
year = {2026},
month = oct,
doi = {10.26434/chemrxiv.15009929/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009929/v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Wavelength-Sequential Generation of Multi-Emissive Fluorophores for Site-Selective Fluorescence Nanoscopy
AU - Saugat Mondal
AU - Tapas Bera
AU - Debjit Maity
AU - Sayan Paul
AU - Swapnamoy Sahoo
AU - Bhaskar Rao Pinninti
AU - Mahitosh Mandal
AU - Hannes A. Houck
AU - N. D. Pradeep Singh
PY - 2026
DA - 2026/10/05
PB - ChemRxiv
DO - 10.26434/chemrxiv.15009929/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009929/v1
AB - Photoactivatable fluorophores are indispensable tools for super-resolution microscopy, but their performance is often hindered by the need for caging group chemistry, ultraviolet activation, interfering triplet-state photochemistry, and most importantly resulting in a single emissive scaffold. Here, we report a hydroacridine-based fluorophore that addresses the often encountered limitations by providing caging-group-free activation using visible light. A heteroatom-engineering strategy enables rapid singlet-state proton-coupled electron transfer, converting a blue-emissive precursor into two highly fluorescent acridinium states with distinct green (ΦF ≈ 0.80) and red (ΦF ≈ 0.78) emission by a two-step photoconversion using blue and green light sequentially. The process operates efficiently under physiological conditions without involving triplet biradical intermediates, enhancing compatibility with live-cell imaging. The fluorophore architecture further supports organelle-targeted imaging and photoinduced intracellular redistribution, enabling multicolour visualization from a single molecular scaffold, serving both spatial and temporal control over the photoactivation. These findings provide new strategies for heteroatom-controlled visible-light photoactivation and fluorophore design, providing a new platform for next-generation super-resolution imaging.
ER -