Evanescent scattering imaging of single protein binding kinetics and DNA conformation changes

Pengfei Zhang, Lei Zhou, Rui Wang, Xinyu Zhou, Jiapei Jiang, Zijian Wan and Shaopeng Wang ()
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Pengfei Zhang: Arizona State University
Lei Zhou: Arizona State University
Rui Wang: Arizona State University
Xinyu Zhou: Arizona State University
Jiapei Jiang: Arizona State University
Zijian Wan: Arizona State University
Shaopeng Wang: Arizona State University

Nature Communications, 2022, vol. 13, issue 1, 1-10

Abstract: Abstract Evanescent illumination has been widely used to detect single biological macromolecules because it can notably enhance light-analyte interaction. However, the current evanescent single-molecule detection system usually requires specially designed microspheres or nanomaterials. Here we show that single protein detection and imaging can be realized on a plain glass surface by imaging the interference between the evanescent lights scattered by the single proteins and by the natural roughness of the cover glass. This allows us to quantify the sizes of single proteins, characterize the protein–antibody interactions at the single-molecule level, and analyze the heterogeneity of single protein binding behaviors. In addition, owing to the exponential distribution of evanescent field intensity, the evanescent imaging system can track the analyte axial movement with high resolution, which can be used to analyze the DNA conformation changes, providing one solution for detecting small molecules, such as microRNA. This work demonstrates a label-free single protein imaging method with ordinary consumables and may pave a road for detecting small biological molecules.

Date: 2022
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DOI: 10.1038/s41467-022-30046-8

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