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• W2004673661 abstract "Dense coverage of DNA by proteins is a ubiquitous feature of cellular processes such as DNA replication, transcription, repair, and compaction. We present a single-molecule manipulation and visualization approach capable of studying individual DNA-protein interactions in the presence of a high density of proteins on the DNA and in solution. This approach combines optical tweezers with multicolor confocal fluorescence microscopy and STED nanoscopy. We demonstrate visualization of proteins on DNA with a spatial resolution of 48 nm, about six-fold better than with traditional wide-field microscopy. The resolution enhancement along the direction of the DNA can be seen in the figure below (scale bar 1μm). Two proteins positioned within the diffraction limit can clearly be distinguished in the STED image. In combination with fast confocal line scanning, STED allows real-time imaging of DNA-protein dynamics with a temporal resolution better than 50 ms. The individual trajectories of proteins translocating on DNA can be distinguished at high protein density and tracked with enhanced localization precision. This unique multimodal approach allowed us to visualize, in real time, the assembly of dense nucleoprotein filaments on DNA with unprecedented spatial resolution.View Large Image | View Hi-Res Image | Download PowerPoint Slide" @default.
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• W2004673661 date "2013-01-01" @default.
• W2004673661 modified "2023-09-26" @default.
• W2004673661 title "STED Nanoscopy Combined with Optical Tweezers Reveals Spatial Dynamics of Proteins on Densely Covered DNA" @default.
• W2004673661 doi "https://doi.org/10.1016/j.bpj.2012.11.1190" @default.
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