Single-image measurements of monochromatic subdiffraction dimolecular separations
Abstract
Measuring subdiffraction separations between single fluorescent particles is important for biological, nano-, and medical-technology studies. Major challenges include (i) measuring changing molecular separations with high temporal resolution while (ii) using identical fluorescent labels. Here we report a method that measures subdiffraction separations between two identical fluorophores by using a single image of milliseconds exposure time and a standard single-molecule fluorescent imaging setup. The fluorophores do not need to be bleached and the separations can be measured down to 40 nm with nanometer precision. The method is called single-molecule image deconvolution -- SMID, and in this article it measures the standard deviation (SD) of Gaussian-approximated combined fluorescent intensity profiles of the two subdiffraction-separated fluorophores. This study enables measurements of (i) subdiffraction dimolecular separations using a single image, lifting the temporal resolution of seconds to milliseconds, while (ii) using identical fluorophores. The single-image nature of this dimer separation study makes it a single-image molecular analysis (SIMA) study.
Cite
@article{arxiv.1002.1980,
title = {Single-image measurements of monochromatic subdiffraction dimolecular separations},
author = {Shawn H. DeCenzo and Michael C. DeSantis and Y. M. Wang},
journal= {arXiv preprint arXiv:1002.1980},
year = {2015}
}
Comments
16 pages, 5 figures