Electrochemically modulated single-molecule localization microscopy for in vitro imaging cytoskeletal protein structures

Lei, Chenghong; Hu, Dehong

doi:10.1515/nanoph-2024-0559

Electrochemically modulated single-molecule localization microscopy for in vitro imaging cytoskeletal protein structures

Journal Article · Sun Feb 09 23:00:00 EST 2025 · Nanophotonics (Online)

DOI:https://doi.org/10.1515/nanoph-2024-0559· OSTI ID:2584257

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Guilin Univ. of Technology (China)
Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

A new concept of electrochemically modulated single-molecule localization super-resolution imaging is developed. Applications of single-molecule localization super-resolution microscopy have been limited due to insufficient availability of qualified fluorophores with favorable low duty cycles. The key for the new concept is that the “On” state of a redox-active fluorophore with unfavorable high duty cycle could be driven to “Off” state by electrochemical potential modulation and thus become available for single-molecule localization imaging. The new concept was carried out using redox-active cresyl violet with unfavorable high duty cycle as a model fluorophore by synchronizing electrochemical potential scanning with a single-molecule localization microscope. The two cytoskeletal protein structures, the microtubules from porcine brain and the actins from rabbit muscle, were selected as the model target structures for the conceptual imaging in vitro. The super-resolution images of microtubules and actins were obtained from precise single-molecule localizations determined by modulating the On/Off states of single fluorophore molecules on the cytoskeletal proteins via electrochemical potential scanning. Importantly, this method could allow more fluorophores even with unfavorable photophysical properties to become available for a wider and more extensive application of single-molecule localization microscopy.

View Accepted Manuscript (DOE)

Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Natural Science Foundation of China (NSFC); Natural Science Foundation of Guangxi Zhuang Autonomous Region

Grant/Contract Number:: AC05-76RL01830

OSTI ID:: 2584257

Report Number(s):: PNNL-SA--181909

Journal Information:: Nanophotonics (Online), Journal Name: Nanophotonics (Online) Journal Issue: 4 Vol. 14; ISSN 2192-8614

Publisher:: de GruyterCopyright Statement

Country of Publication:: United States

Language:: English

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