RT Journal Article
SR Electronic
T1 Nanoscopic Stoichiometry and Single-Molecule Counting
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 524801
DO 10.1101/524801
A1 Daniel Nino
A1 Daniel Djayakarsana
A1 Joshua N. Milstein
YR 2019
UL http://biorxiv.org/content/early/2019/01/18/524801.abstract
AB Single-molecule localization microscopy (SMLM) has the potential to revolutionize proteomic and genomic analyses by providing information on the number and stoichiometry of proteins or nucleic acids aggregating at spatial scales below the diffraction limit of light. Here we present a method for molecular counting with SMLM built upon the exponentially distributed blinking statistics of photoswitchable fluorophores, with a focus on organic dyes. We provide a practical guide to molecular counting, highlighting many of the challenges and pitfalls, by benchmarking the method on fluorescently labeled, surface mounted DNA origami grids. The accuracy of the results illustrates SMLM’s utility for optical ‘-omics’ analysis.