Scanning Electron Microscopy Imaging of Large DNA Molecules Using a Metal-Free Electro-Stain Composed of DNA-Binding Proteins and Synthetic Polymers.

Publisher: WILEY-VCH Country of Publication: Germany NLM ID: 101664569 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2198-3844 (Electronic) Linking ISSN: 21983844 NLM ISO Abbreviation: Adv Sci (Weinh) Subsets: MEDLINE

Original Publication: Weinheim : WILEY-VCH, [2014]-

Microscopy, Electron, Scanning*/methods ; DNA*/chemistry ; DNA*/metabolism ; Polymers*/chemistry ; DNA-Binding Proteins*/metabolism ; DNA-Binding Proteins*/ultrastructure ; DNA-Binding Proteins*/chemistry; Quantum Dots/chemistry

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.
(© 2024 The Authors. Advanced Science published by Wiley‐VCH GmbH.)

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SRFC-MA2101-07 Samsung Science and Technology Foundation

Keywords: DNA binding protein; SEM DNA imaging; SEM optical DNA mapping; polyvinylpyrrolidone; quantum dot labeling

9007-49-2 (DNA)
0 (Polymers)
0 (DNA-Binding Proteins)

Date Created: 20240505 Date Completed: 20240724 Latest Revision: 20240726

20240726

PMC11267313

10.1002/advs.202309702

38704672