Scanning Electron Microscopy Imaging of Large DNA Molecules Using a Metal-Free Electro-Stain Composed of DNA-Binding Proteins and Synthetic Polymers
  • Noh, Chanyoung
  • Kang, Yoonjung
  • Heo, Sujung
  • Kim, Taesoo
  • Kim, Hayeon
  • ... Lim, Kwang-il
  • 외 5명
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초록

This paper presents the first scanning electron microscopy (SEM)-based DNA imaging in biological samples. This novel approach incorporates a metal-free electro-stain reagent, formulated by combining DNA-binding proteins and synthetic polymers to enhance the visibility of 2-nm-thick DNA under SEM. Notably, DNA molecules stain with proteins and polymers appear as dark lines under SEM. The resulting DNA images exhibit a thickness of 15.0±4.0 nm. As SEM is the primary platform, it integrates seamlessly with various chemically functionalized large surfaces with the aid of microfluidic devices. The approach allows high-resolution imaging of various DNA structures including linear, circular, single-stranded DNA and RNA, originating from nuclear and mitochondrial genomes. Furthermore, quantum dots are successfully visualized as bright labels that are sequence-specifically incorporated into DNA molecules, which highlights the potential for SEM-based optical DNA mapping. In conclusion, DNA imaging using SEM with the novel electro-stain offers electron microscopic resolution with the ease of optical microscopy.

키워드

DNA binding proteinpolyvinylpyrrolidonequantum dot labelingSEM DNA imagingSEM optical DNA mappingSINGLE-MOLECULELOCALIZATION MICROSCOPYDIRECT VISUALIZATIONSTRUCTURAL VARIATIONTRANSCRIPTIONHELIX
제목
Scanning Electron Microscopy Imaging of Large DNA Molecules Using a Metal-Free Electro-Stain Composed of DNA-Binding Proteins and Synthetic Polymers
저자
Noh, ChanyoungKang, YoonjungHeo, SujungKim, TaesooKim, HayeonChang, JunhyuckSundharbaabu, Priyannth RamasamiShim, SangheeLim, Kwang-ilLee, Jung HeonJo, Kyubong
DOI
10.1002/advs.202309702
발행일
2024-07
유형
Article; Early Access
저널명
Advanced Science
11
28