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A wavelength-induced frequency filtering method for fluorescent nanosensors in vivo

Abstract

Fluorescent nanosensors hold the potential to revolutionize life sciences and medicine. However, their adaptation and translation into the in vivo environment is fundamentally hampered by unfavourable tissue scattering and intrinsic autofluorescence. Here we develop wavelength-induced frequency filtering (WIFF) whereby the fluorescence excitation wavelength is modulated across the absorption peak of a nanosensor, allowing the emission signal to be separated from the autofluorescence background, increasing the desired signal relative to noise, and internally referencing it to protect against artefacts. Using highly scattering phantom tissues, an SKH1-E mouse model and other complex tissue types, we show that WIFF improves the nanosensor signal-to-noise ratio across the visible and near-infrared spectra up to 52-fold. This improvement enables the ability to track fluorescent carbon nanotube sensor responses to riboflavin, ascorbic acid, hydrogen peroxide and a chemotherapeutic drug metabolite for depths up to 5.5 ± 0.1 cm when excited at 730 nm and emitting between 1,100 and 1,300 nm, even allowing the monitoring of riboflavin diffusion in thick tissue. As an application, nanosensors aided by WIFF detect the chemotherapeutic activity of temozolomide transcranially at 2.4 ± 0.1 cm through the porcine brain without the use of fibre optic or cranial window insertion. The ability of nanosensors to monitor previously inaccessible in vivo environments will be important for life-sciences research, therapeutics and medical diagnostics.

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Fig. 1: Challenges for deep-tissue sensing.
Fig. 2: Principle of WIFF.
Fig. 3: WIFF performance in a complex tissue.
Fig. 4: Elucidating the effect of autofluorescence on WIFF and deep-tissue detection.
Fig. 5: In vivo sensing.
Fig. 6: Transcranial dynamics of the chemotherapeutic metabolite AIC in porcine brain.

Data availability

Source data are available via Zenodo at https://doi.org/10.5281/zenodo.6049452. The data that support the findings of this study are available in the paper and Supplementary Information. All other data are available from the corresponding author upon reasonable request.

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Acknowledgements

The research is supported by the Koch Institute for Integrative Cancer Research at MIT and the Bridge Project Program. V.B.K. is supported by The Swiss National Science Foundation (project nos. P2ELP3_162149 and P300P2_174469). D.K. is supported by the Grant-in-Aid for JSPS Fellows (JSPS KAKENHI grant no. 15J07423) and Encouragement of Young Scientists (B) (JSPS KAKENHI grant no. JP16K17485) from the Japan Society for the Promotion of Science. X.J. is supported by the King Abdullah University of Science & Technology (OSR-2015 Sensors 2707). G.B. acknowledges support from the Zuckerman STEM Leadership Program and the Israeli Science Foundation (grant no. 456/18). F.T.N. is supported by the Arnold O. Beckman Postdoctoral Fellowship from the Arnold and Mabel Beckman Foundation. V.B.K. acknowledges helpful discussions with J. Yang.

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V.B.K. and M.S.S. conceived the idea and planned the experiments with the assistance of N.A.B., X.J. and F.T.N. V.B.K. developed the experimental setup, performed the in vitro experiments and analysed the data with the assistance of D.K., F.T.N., M.A.L., G.B. and J.D. F.T.N. and V.B.K. performed the tissue autofluorescence studies. N.A.B. and X.J. performed the ex vivo and in vivo experiments with the assistance of V.B.K. V.B.K. and M.S.S. wrote the manuscript with inputs from all the authors. All the authors contributed to discussions regarding the research.

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Correspondence to Michael S. Strano.

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Nature Nanotechnology thanks Yun-Sheng Chen and Fan Zhang for their contribution to the peer review of this work.

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Koman, V.B., Bakh, N.A., Jin, X. et al. A wavelength-induced frequency filtering method for fluorescent nanosensors in vivo. Nat. Nanotechnol. 17, 643–652 (2022). https://doi.org/10.1038/s41565-022-01136-x

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