{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Shaw TR"],"funding":["National Institutes of Health","NIGMS NIH HHS","National Science Foundation"],"pagination":["2906-2920"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9388596"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["121(15)"],"pubmed_abstract":["Single-molecule localization microscopy (SMLM) permits the visualization of cellular structures an order of magnitude smaller than the diffraction limit of visible light, and an accurate, objective evaluation of the resolution of an SMLM data set is an essential aspect of the image processing and analysis pipeline. Here, we present a simple method to estimate the localization spread function (LSF) of a static SMLM data set directly from acquired localizations, exploiting the correlated dynamics of individual emitters and properties of the pair autocorrelation function evaluated in both time and space. The method is demonstrated on simulated localizations, DNA origami rulers, and cellular structures labeled by dye-conjugated antibodies, DNA-PAINT, or fluorescent fusion proteins. We show that experimentally obtained images have LSFs that are broader than expected from the localization precision alone, due to additional uncertainty accrued when localizing molecules imaged over time."],"journal":["Biophysical journal"],"pubmed_title":["Estimating the localization spread function of static single-molecule localization microscopy images."],"pmcid":["PMC9388596"],"funding_grant_id":["R01 GM129347","K12 GM111725","GM129347","GM110052","R01 GM110052","MCB1552439"],"pubmed_authors":["Kim S","Fazekas FJ","Flanagan-Natoli JC","Sumrall ER","Veatch SL","Shaw TR"],"additional_accession":[]},"is_claimable":false,"name":"Estimating the localization spread function of static single-molecule localization microscopy images.","description":"Single-molecule localization microscopy (SMLM) permits the visualization of cellular structures an order of magnitude smaller than the diffraction limit of visible light, and an accurate, objective evaluation of the resolution of an SMLM data set is an essential aspect of the image processing and analysis pipeline. Here, we present a simple method to estimate the localization spread function (LSF) of a static SMLM data set directly from acquired localizations, exploiting the correlated dynamics of individual emitters and properties of the pair autocorrelation function evaluated in both time and space. The method is demonstrated on simulated localizations, DNA origami rulers, and cellular structures labeled by dye-conjugated antibodies, DNA-PAINT, or fluorescent fusion proteins. We show that experimentally obtained images have LSFs that are broader than expected from the localization precision alone, due to additional uncertainty accrued when localizing molecules imaged over time.","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 Aug","modification":"2026-06-04T04:44:32.918Z","creation":"2026-05-05T03:12:25.561Z"},"accession":"S-EPMC9388596","cross_references":{"pubmed":["35787472"],"doi":["10.1016/j.bpj.2022.06.036"]}}