Photobleaching of YOYO-1 in super-resolution single DNA fluorescence imaging

Joseph R Pyle, Jixin Chen
2017 Beilstein Journal of Nanotechnology  
Super-resolution imaging of single DNA molecules via point accumulation for imaging in nanoscale topography (PAINT) has great potential to visualize fine DNA structures with nanometer resolution. In a typical PAINT video acquisition, dye molecules (YOYO-1) in solution sparsely bind to the target surfaces (DNA) whose locations can be mathematically determined by fitting their fluorescent point spread function. Many YOYO-1 molecules intercalate into DNA and remain there during imaging, and most
more » ... them have to be temporarily or permanently fluorescently bleached, often stochastically, to allow for the visualization of a few fluorescent events per DNA per frame of the video. Thus, controlling the fluorescence on-off rate is important in PAINT. In this paper, we study the photobleaching of YOYO-1 and its correlation with the quality of the PAINT images. At a low excitation laser power density, the photobleaching of YOYO-1 is too slow and a minimum required power density was identified, which can be theoretically predicted with the proposed method in this report. 2296 Supporting Information Additional experimental information. Supporting Information File 1 Laser spot size, DNA length, PSF, and photocount histograms. [http://www.beilstein-journals.org/bjnano/content/ supplementary/2190-4286-8-229-S1.pdf]
doi:10.3762/bjnano.8.229 pmid:29181286 pmcid:PMC5687005 fatcat:zfo6ptazgvgh7hg6rb5sbhnsji