Investigating Single-Molecule Fluorescence Spectral Heterogeneity of Rhodamines Using High-Throughput Single-Molecule Spectroscopy

Zhang, Yang; Zhang, Yu; Song, Ki-Hee; Lin, Wei; Sun, Cheng; Schatz, George C.; Zhang, Hao F.

doi:10.1021/acs.jpclett.1c00192

Investigating Single-Molecule Fluorescence Spectral Heterogeneity of Rhodamines Using High-Throughput Single-Molecule Spectroscopy

Journal Article · Fri Apr 16 00:00:00 EDT 2021 · Journal of Physical Chemistry Letters

DOI:https://doi.org/10.1021/acs.jpclett.1c00192· OSTI ID:1798266

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Northwestern Univ., Evanston, IL (United States); Northwestern University
Northwestern Univ., Evanston, IL (United States)

We experimentally investigated several intra-molecular coordinate and environmental changes as potential causes of single-molecule fluorescence spectral heterogeneities (smFSH). Here, we developed a high-throughput single-molecule spectroscopy technique to analyze more than 5,000 single-molecule emission spectra from each of nine commonly used fluorophores with different structural rigidities deposited on substrates with different polarities. We observed an unexpected high smFSH from structurally rigid Rhodamine B compared with a structurally flexible Cyanine dye - Alexa Fluor 647. Based on experimentally measured smFSH, we ruled out the system’s noise uncertainty, single-molecule spectral diffusion, and environmental polarity as the primary causes of the high smFSH. We found that the rotational flexibility of N, N-dialkylated groups contributed to the smFSH. With the high smFSH observed in structurally more rigid model fluorophores, we speculated that other intra-molecular coordinate and environmental changes might also contribute to the high smFSH in Rhodamines.

View Accepted Manuscript (DOE)

Research Organization:: Northwestern Univ., Evanston, IL (United States)

Sponsoring Organization:: NIH; NSF; USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division

Grant/Contract Number:: SC0004752

OSTI ID:: 1798266

Journal Information:: Journal of Physical Chemistry Letters, Journal Name: Journal of Physical Chemistry Letters Journal Issue: 16 Vol. 12; ISSN 1948-7185

Publisher:: American Chemical SocietyCopyright Statement

Country of Publication:: United States

Language:: English

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