Molecular specificity of nucleic acids via intrinsic stochastic fluorescence under visible light illumination

Ruyi Gong; Luay Almassalha; Vadim Backman

doi:10.1117/12.3001893

13 March 2024 Molecular specificity of nucleic acids via intrinsic stochastic fluorescence under visible light illumination

Ruyi Gong, Luay Almassalha, Vadim Backman

Author Affiliations +

Proceedings Volume PC12849, Single Molecule Spectroscopy and Superresolution Imaging XVII; PC128490A (2024) https://doi.org/10.1117/12.3001893
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

The development of super-resolution imaging techniques have extended the resolving power to around 10 - 50 nm. However, most current super-resolution imaging techniques need exogenous fluorescent dyes as imaging contrast, whose essential weakness of labeling includes imprecise spatial localization, and perturbation of the sample. In 2017, Dong et al., demonstrated that the intrinsic fluorescence of DNA under visible light excitation has similar photo-switching properties to the organic dyes used in single-molecule localization microscopy. In this paper, we measured the fluorescence spectra of poly-G (guanine) of different lengths (5, 8, 12, 16 base-pair), 20 base-pair single-stranded DNA molecules (poly-A, G, C, T), as well as double-stranded DNA (AT chain, GC chain), under multiple wavelengths. The 20 base-pair AT, GC spectra can be classified with an accuracy of more than 90%, which demonstrates the molecular specificity of the double stranded DNA polymers via its intrinsic fluorescence. Our work paves the way for developing spectroscopic intrinsic-contrast localization optical nanoscopy for chromatin study.

Conference Presentation

Citation Download Citation

Ruyi Gong, Luay Almassalha, and Vadim Backman "Molecular specificity of nucleic acids via intrinsic stochastic fluorescence under visible light illumination", Proc. SPIE PC12849, Single Molecule Spectroscopy and Superresolution Imaging XVII, PC128490A (13 March 2024); https://doi.org/10.1117/12.3001893

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