title: Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging.
abstract: One approach to super-resolution fluorescence imaging uses sequential activation and localization of individual fluorophores to achieve high spatial resolution. Essential to this technique is the choice of fluorescent probes; the properties of the probes, including photons per switching event, on-off duty cycle, photostability and number of switching cycles, largely dictate the quality of super-resolution images. Although many probes have been reported, a systematic characterization of the properties of these probes and their impact on super-resolution image quality has been described in only a few cases. Here we quantitatively characterized the switching properties of 26 organic dyes and directly related these properties to the quality of super-resolution images. This analysis provides guidelines for characterization of super-resolution probes and a resource for selecting probes based on performance. Our evaluation identified several photoswitchable dyes with good to excellent performance in four independent spectral ranges, with which we demonstrated low-cross-talk, four-color super-resolution imaging.
authors: Graduate program in Biophysics, Harvard University, Cambridge, Massachusetts, USA.
authors: Graham T
authors: GT
authors: Dempsey
authors: Joshua C
authors: JC
authors: Vaughan
authors: Kok Hao
authors: KH
authors: Chen
authors: Mark
authors: M
authors: Bates
authors: Xiaowei
authors: X
authors: Zhuang
mesh_terms: D002021:Buffers; D003116:Color; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis*; D008027:Light; D008163:Luminescent Measurements; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism; D057054:Molecular Imaging / Q000379:methods*; D015335:Molecular Probes / Q000032:analysis / Q000737:chemistry; D010316:Particle Size; D017785:Photons
chemical_list: D002021:Buffers; D005456:Fluorescent Dyes; D015335:Molecular Probes