Single Protein Detection and Imaging with Evanescent Scattering Microscopy

Pengfei Zhang, Lei Zhou, Rui Wang, Xinyu Zhou, Jiapei Jiang, Zijian Wan, Shaopeng Wang

Research output: Contribution to journalArticlepeer-review


Single-molecule measurements provide statistical distributions of molecular properties, in addition to the ensemble averages. Evanescent detection approaches have been widely used for single-molecule detection because the evanescent field can significantly enhance the light-analyte interaction and reduce the background noise. However, current evanescent single-molecule detection systems mostly require specially designed sensing components. Here, we show that single proteins can be imaged on a plain cover glass surface by detecting the evanescent waves scattered by the target molecules. This allows us to quantify the protein–antibody interactions at the single-molecule level. This protocol describes a label-free single-molecule imaging approach with conventional consumables and may pave the road for detecting single molecules with commercial optical microscopy.

Original languageEnglish (US)
Article numbere4530
Issue number20
StatePublished - Oct 20 2022


  • Cover glass
  • Evanescent field
  • Scattering microscopy
  • Single protein
  • Single-molecule imaging
  • Total internal reflection

ASJC Scopus subject areas

  • General Neuroscience
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology
  • Plant Science


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