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<p>DNA-Point Accumulation for Imaging in Nanoscale Topography (DNA-PAINT) is a versatile super-resolution technique that relies on the predictable binding kinetics between fluorescent imager strands and docking strands attached to target proteins. This makes DNA-PAINT particularly suitable for multiplexing and quantitative applications, but its performance is often limited by spurious signals from non-specific binding of imager strands. Here we describe a method to remove these non-specific binding events using a statistical test to distinguish between DNA-specific and non-specific interactions. To demonstrate the method, we imaged mosaic epithelial tissues in <i>Drosophila melanogaster</i> egg chambers and showed that <b>&gt;</b>90% of non-specific and otherwise indistinguishable signal in the super-resolved images can be removed. This denoising improves the quality of DNA-PAINT super-resolved images and is essential for accurate measurements of spatial relationships and protein quantification.</p>

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Utilizing the predictable binding kinetics of DNA-PAINT to denoise super-resolution images

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月5日

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  • George Sirinakis,
  • Edward S. Allgeyer,
  • Jennifer H. Richens,
  • Jeanne Lefévère-Laoide,
  • Ewa K. Paluch,
  • Daniel St Johnston

摘要

DNA-Point Accumulation for Imaging in Nanoscale Topography (DNA-PAINT) is a versatile super-resolution technique that relies on the predictable binding kinetics between fluorescent imager strands and docking strands attached to target proteins. This makes DNA-PAINT particularly suitable for multiplexing and quantitative applications, but its performance is often limited by spurious signals from non-specific binding of imager strands. Here we describe a method to remove these non-specific binding events using a statistical test to distinguish between DNA-specific and non-specific interactions. To demonstrate the method, we imaged mosaic epithelial tissues in Drosophila melanogaster egg chambers and showed that >90% of non-specific and otherwise indistinguishable signal in the super-resolved images can be removed. This denoising improves the quality of DNA-PAINT super-resolved images and is essential for accurate measurements of spatial relationships and protein quantification.