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

Author

Listed:
  • Pengfei Zhang

    (Arizona State University)

  • Lei Zhou

    (Arizona State University)

  • Rui Wang

    (Arizona State University)

  • Xinyu Zhou

    (Arizona State University
    Arizona State University)

  • Jiapei Jiang

    (Arizona State University
    Arizona State University)

  • Zijian Wan

    (Arizona State University
    Arizona State University)

  • Shaopeng Wang

    (Arizona State University
    Arizona State University)

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.

Suggested Citation

  • Pengfei Zhang & Lei Zhou & Rui Wang & Xinyu Zhou & Jiapei Jiang & Zijian Wan & Shaopeng Wang, 2022. "Evanescent scattering imaging of single protein binding kinetics and DNA conformation changes," Nature Communications, Nature, vol. 13(1), pages 1-10, December.
  • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-30046-8
    DOI: 10.1038/s41467-022-30046-8
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    References listed on IDEAS

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    1. Amy E. Goodling & Sara Nagelberg & Bryan Kaehr & Caleb H. Meredith & Seong Ik Cheon & Ashley P. Saunders & Mathias Kolle & Lauren D. Zarzar, 2019. "Colouration by total internal reflection and interference at microscale concave interfaces," Nature, Nature, vol. 566(7745), pages 523-527, February.
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    3. Guangzhong Ma & Zijian Wan & Yunze Yang & Pengfei Zhang & Shaopeng Wang & Nongjian Tao, 2020. "Optical imaging of single-protein size, charge, mobility, and binding," Nature Communications, Nature, vol. 11(1), pages 1-11, December.
    4. Marek Piliarik & Vahid Sandoghdar, 2014. "Direct optical sensing of single unlabelled proteins and super-resolution imaging of their binding sites," Nature Communications, Nature, vol. 5(1), pages 1-8, December.
    5. Kaijie Ma & Yunong Zhang & Le Liu & Jingyu Xi & Xinping Qiu & Tian Guan & Yonghong He, 2019. "In situ mapping of activity distribution and oxygen evolution reaction in vanadium flow batteries," Nature Communications, Nature, vol. 10(1), pages 1-11, December.
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