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Single-molecule nucleic acid interactions monitored on a label-free microcavity biosensor platform

Subjects

Abstract

Biosensing relies on the detection of molecules and their specific interactions. It is therefore highly desirable to develop transducers exhibiting ultimate detection limits. Microcavities are an exemplary candidate technology for demonstrating such a capability in the optical domain and in a label-free fashion. Additional sensitivity gains, achievable by exploiting plasmon resonances, promise biosensing down to the single-molecule level. Here, we introduce a biosensing platform using optical microcavity-based sensors that exhibits single-molecule sensitivity and is selective to specific single binding events. Whispering gallery modes in glass microspheres are used to leverage plasmonic enhancements in gold nanorods for the specific detection of nucleic acid hybridization, down to single 8-mer oligonucleotides. Detection of single intercalating small molecules confirms the observation of single-molecule hybridization. Matched and mismatched strands are discriminated by their interaction kinetics. Our platform allows us to monitor specific molecular interactions transiently, hence mitigating the need for high binding affinity and avoiding permanent binding of target molecules to the receptors. Sensor lifetime is therefore increased, allowing interaction kinetics to be statistically analysed.

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Figure 1: Experimental set-up of whispering-gallery-mode sensing platform.
Figure 2: Detection of nanorods in the transient and binding regimes.
Figure 3: Transient nucleic acid interactions monitored with transverse electric and transverse magnetic whispering gallery modes simultaneously.
Figure 4: Discrimination of fully complementary and single-base-mismatched DNA strands by monitoring interaction kinetics for different sodium concentrations.
Figure 5: Detection of short oligonucleotides and small intercalating molecules.

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Acknowledgements

The authors acknowledge financial support for this work from the Max Planck Society (M.D.B. and F.V.) and the Alexander von Humboldt Foundation (M.R.F.).

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F.V. and M.D.B. conceived and planned the experiments. M.D.B. conducted experimental work and data analysis. M.R.F. performed numerical and theoretical analysis. F.V., M.R.F. and M.D.B. wrote the manuscript.

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Correspondence to Frank Vollmer.

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The authors declare no competing financial interests.

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Baaske, M., Foreman, M. & Vollmer, F. Single-molecule nucleic acid interactions monitored on a label-free microcavity biosensor platform. Nature Nanotech 9, 933–939 (2014). https://doi.org/10.1038/nnano.2014.180

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