{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T06:19:12Z","timestamp":1772173152510,"version":"3.50.1"},"update-to":[{"DOI":"10.1371\/journal.pcbi.1010765","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2023,1,9]],"date-time":"2023-01-09T00:00:00Z","timestamp":1673222400000}}],"reference-count":43,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2022,12,27]],"date-time":"2022-12-27T00:00:00Z","timestamp":1672099200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000275","name":"Leverhulme Trust","doi-asserted-by":"publisher","award":["RPG-2017-349"],"award-info":[{"award-number":["RPG-2017-349"]}],"id":[{"id":"10.13039\/501100000275","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000275","name":"Leverhulme Trust","doi-asserted-by":"publisher","award":["RPG-2017-349"],"award-info":[{"award-number":["RPG-2017-349"]}],"id":[{"id":"10.13039\/501100000275","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["106151\/Z\/14\/Z"],"award-info":[{"award-number":["106151\/Z\/14\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Royal Society Wolfson Research Merit Award","award":["WM150020"],"award-info":[{"award-number":["WM150020"]}]},{"name":"Medical Research Council Doctoral Training Partnership grant","award":["MR\/J003964\/1"],"award-info":[{"award-number":["MR\/J003964\/1"]}]}],"content-domain":{"domain":["www.ploscompbiol.org"],"crossmark-restriction":false},"short-container-title":["PLoS Comput Biol"],"abstract":"<jats:p>Gaussian spot fitting methods have significantly extended the spatial range where fluorescent microscopy can be used, with recent techniques approaching nanometre (nm) resolutions. However, small inter-fluorophore distances are systematically over-estimated for typical molecular scales. This bias can be corrected computationally, but current algorithms are limited to correcting distances between pairs of fluorophores. Here we present a flexible Bayesian computational approach that infers the distances and angles between multiple fluorophores and has several advantages over these previous methods. Specifically it improves confidence intervals for small lengths, estimates measurement errors of each fluorophore individually and infers the correlations between polygon lengths. The latter is essential for determining the full multi-fluorophore 3D architecture. We further developed the algorithm to infer the mixture composition of a heterogeneous population of multiple polygon states. We use our algorithm to analyse the 3D architecture of the human kinetochore, a macro-molecular complex that is essential for high fidelity chromosome segregation during cell division. Using triple fluorophore image data we unravel the mixture of kinetochore states during human mitosis, inferring the conformation of microtubule attached and unattached kinetochores and their proportions across mitosis. We demonstrate that the attachment conformation correlates with intersister tension and sister alignment to the metaphase plate.<\/jats:p>","DOI":"10.1371\/journal.pcbi.1010765","type":"journal-article","created":{"date-parts":[[2022,12,27]],"date-time":"2022-12-27T13:46:07Z","timestamp":1672148767000},"page":"e1010765","update-policy":"https:\/\/doi.org\/10.1371\/journal.pcbi.corrections_policy","source":"Crossref","is-referenced-by-count":6,"title":["Bayesian inference of multi-point macromolecular architecture mixtures at nanometre resolution"],"prefix":"10.1371","volume":"18","author":[{"given":"Peter A.","family":"Embacher","sequence":"first","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0461-5374","authenticated-orcid":true,"given":"Tsvetelina E.","family":"Germanova","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4125-4632","authenticated-orcid":true,"given":"Emanuele","family":"Roscioli","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6808-0711","authenticated-orcid":true,"given":"Andrew D.","family":"McAinsh","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4632-1550","authenticated-orcid":true,"given":"Nigel J.","family":"Burroughs","sequence":"additional","affiliation":[]}],"member":"340","published-online":{"date-parts":[[2022,12,27]]},"reference":[{"key":"pcbi.1010765.ref001","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1038\/nmeth.2843","article-title":"Precisely and accurately localizing single emitters in fluorescence microscopy","volume":"11","author":"H Deschout","year":"2014","journal-title":"Nature Methods"},{"key":"pcbi.1010765.ref002","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1038\/nature09163","article-title":"Subnanometre single-molecule localization, registration and distance measurements","volume":"466","author":"A Pertsinidis","year":"2010","journal-title":"Nature"},{"key":"pcbi.1010765.ref003","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1021\/ar040136s","article-title":"Fluorescence Imaging with One Nanometer Accuracy: Application to Molecular Motors","volume":"38","author":"A Yildiz","year":"2005","journal-title":"Acc. Chem. Res."},{"issue":"5","key":"pcbi.1010765.ref004","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1038\/nmeth.1447","article-title":"Optimized localization analysis for single-molecule tracking and super-resolution microscopy","volume":"7","author":"KI Mortensen","year":"2010","journal-title":"Nature Methods"},{"key":"pcbi.1010765.ref005","doi-asserted-by":"crossref","first-page":"2775","DOI":"10.1016\/S0006-3495(02)75618-X","article-title":"Precise Nanometer Localization Analysis for Individual Fluorescent Probes","volume":"82","author":"RE Thompson","year":"2002","journal-title":"Biophysical Journal"},{"key":"pcbi.1010765.ref006","doi-asserted-by":"crossref","first-page":"668","DOI":"10.1529\/biophysj.105.065599","article-title":"A Non-Gaussian Distribution Quantifies Distances Measured with Fluorescence Localization Techniques","volume":"90","author":"LS Churchman","year":"2006","journal-title":"Biophysical Journal"},{"key":"pcbi.1010765.ref007","doi-asserted-by":"crossref","DOI":"10.1007\/978-3-030-40183-2","volume-title":"Probability Theory and Stochastic Processes","author":"P Br\u00e9maud","year":"2020"},{"issue":"5","key":"pcbi.1010765.ref008","doi-asserted-by":"crossref","first-page":"1419","DOI":"10.1073\/pnas.0409487102","article-title":"Single molecule high-resolution colocalization of Cy3 and Cy5 attached to macromolecules measures intramolecular distances through time","volume":"102","author":"LS Churchman","year":"2005","journal-title":"PNAS"},{"issue":"4","key":"pcbi.1010765.ref009","doi-asserted-by":"crossref","first-page":"100774","DOI":"10.1016\/j.xpro.2021.100774","article-title":"Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells","volume":"2","author":"TE Germanova","year":"2021","journal-title":"STAR Protocols"},{"issue":"10","key":"pcbi.1010765.ref010","doi-asserted-by":"crossref","first-page":"4275","DOI":"10.1073\/pnas.1815826116","article-title":"Nanometer-accuracy distance measurements between fluorophores at the single-molecule level","volume":"116","author":"S Niekamp","year":"2019","journal-title":"PNAS"},{"key":"pcbi.1010765.ref011","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1021\/acs.nanolett.0c03332","article-title":"Nanoscale Pattern Extraction from Relative Positions of Sparse 3D Localizations","volume":"21","author":"AP Curd","year":"2021","journal-title":"Nano Letters"},{"issue":"4","key":"pcbi.1010765.ref012","doi-asserted-by":"crossref","DOI":"10.1016\/j.celrep.2020.107535","article-title":"Ensemble-level organization of human kinetochores and evidence for distinct tension and attachment sensors","volume":"31","author":"E Roscioli","year":"2020","journal-title":"Cell Reports"},{"issue":"5793","key":"pcbi.1010765.ref013","doi-asserted-by":"crossref","first-page":"1642","DOI":"10.1126\/science.1127344","article-title":"Imaging Intracellular Fluorescent Proteins at Nanometer Resolution","volume":"313","author":"E Betzig","year":"2006","journal-title":"Science"},{"key":"pcbi.1010765.ref014","doi-asserted-by":"crossref","first-page":"13890","DOI":"10.1021\/acs.chemrev.7b00218","article-title":"Super-Resolution Structured Illumination Microscopy","volume":"117","author":"R Heintzmann","year":"2017","journal-title":"Chemical Reviews"},{"key":"pcbi.1010765.ref015","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.biocel.2018.05.014","article-title":"SRRF: Universal live-cell super-resolution microscopy","volume":"101","author":"S Culley","year":"2018","journal-title":"The International Journal of Biochemistry & Cell Biology"},{"key":"pcbi.1010765.ref016","doi-asserted-by":"crossref","first-page":"014003","DOI":"10.1088\/2050-6120\/3\/1\/014003","article-title":"Resolution improvement by 3D particle averaging in localization microscopy","volume":"3","author":"J Broeken","year":"2015","journal-title":"Methods Appl. Fluoresc"},{"key":"pcbi.1010765.ref017","doi-asserted-by":"crossref","first-page":"2847","DOI":"10.1038\/s41467-021-22006-5","article-title":"3D particle averaging and detection of macromolecular symmetry in localization microscopy","volume":"12","author":"H Heydarian","year":"2021","journal-title":"Nature Communications"},{"key":"pcbi.1010765.ref018","doi-asserted-by":"crossref","first-page":"2379","DOI":"10.1038\/s41467-019-10007-4","article-title":"A tessellation-based colocalization analysis approach for single-molecule localization microscopy","volume":"10","author":"F Levet","year":"2019","journal-title":"Nature Communications"},{"key":"pcbi.1010765.ref019","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00418-011-0880-5","article-title":"Coordinate-based colocalization analysis of single-molecule localization microscopy data","volume":"137","author":"S Malkusch","year":"2012","journal-title":"Histochemistry and Cell Biology"},{"issue":"11","key":"pcbi.1010765.ref020","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1038\/nmeth.1704","article-title":"Probing protein heterogeneity in the plasma membrane using PALM and pair correlation analysis","volume":"8","author":"P Sengupta","year":"2011","journal-title":"Nature Methods"},{"key":"pcbi.1010765.ref021","doi-asserted-by":"crossref","first-page":"1387","DOI":"10.1038\/nbt896","article-title":"FRET imaging","volume":"21","author":"EA Jares-Erijman","year":"2003","journal-title":"Nature Biotechnology"},{"key":"pcbi.1010765.ref022","volume-title":"Bayesian Data Analysis","author":"A Gelman","year":"2004","edition":"2"},{"key":"pcbi.1010765.ref023","volume-title":"Markov chain Monte Carlo in practice","author":"WR Gilks","year":"1996"},{"key":"pcbi.1010765.ref024","doi-asserted-by":"crossref","DOI":"10.1007\/978-0-387-21811-3","volume-title":"Spatial Statistics and Computational Methods","author":"J M\u00f8ller","year":"2003"},{"issue":"5","key":"pcbi.1010765.ref025","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1083\/jcb.201211113","article-title":"The functions and consequences of force at kinetochores","volume":"200","author":"F Rago","year":"2013","journal-title":"J. Cell Biol."},{"key":"pcbi.1010765.ref026","doi-asserted-by":"crossref","first-page":"906","DOI":"10.1038\/emboj.2011.15","article-title":"A TACC3\/ch-TOG\/clathrin complex stabilises kinetochore fibres by inter-microtubule bridging","volume":"30","author":"DG Booth","year":"2011","journal-title":"EMBO J"},{"issue":"17","key":"pcbi.1010765.ref027","doi-asserted-by":"crossref","first-page":"2551","DOI":"10.1038\/emboj.2009.173","article-title":"The life and miracles of kinetochores","volume":"28","author":"S Santaguida","year":"2009","journal-title":"EMBO Journal"},{"issue":"11","key":"pcbi.1010765.ref028","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1016\/j.cub.2004.05.053","article-title":"Spindle Checkpoint Protein Dynamics at Kinetochores in Living Cells","volume":"14","author":"BJ Howell","year":"2004","journal-title":"Current Biology"},{"issue":"8161","key":"pcbi.1010765.ref029","article-title":"A quantitative description of Ndc80 complex linkage to human kinetochores","volume":"6","author":"A Suzuki","year":"2015","journal-title":"Nature Communications"},{"key":"pcbi.1010765.ref030","doi-asserted-by":"crossref","first-page":"200101","DOI":"10.1098\/rsob.200101","article-title":"Effects of malleable kinetochore morphology on measurements of intrakinetochore tension","volume":"10","author":"F Renda","year":"2020","journal-title":"Open Biol"},{"key":"pcbi.1010765.ref031","doi-asserted-by":"crossref","first-page":"1567","DOI":"10.1083\/jcb.137.7.1567","article-title":"Kinetochore fiber maturation in PtK1 cells and its implications for the mechanisms of chromosome congression and anaphase onset","volume":"137","author":"BF McEwen","year":"1997","journal-title":"J. Cell Biol"},{"issue":"5285","key":"pcbi.1010765.ref032","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1126\/science.274.5285.242","article-title":"Association of Spindle Assembly Checkpoint Component XMAD2 with Unattached Kinetochores","volume":"274","author":"R-H Chen","year":"1996","journal-title":"Science"},{"issue":"6","key":"pcbi.1010765.ref033","doi-asserted-by":"crossref","first-page":"1533","DOI":"10.1083\/jcb.201611104","article-title":"Spindle assembly checkpoint satisfaction occurs via end-on but not lateral attachments under tension","volume":"216","author":"J Kuhn","year":"2017","journal-title":"J. Cell Biol"},{"issue":"5","key":"pcbi.1010765.ref034","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1083\/jcb.200909005","article-title":"Kinetochore alignment within the metaphase plate is regulated by centromere stiffness and microtubule depolymerases","volume":"188","author":"K Jaqaman","year":"2010","journal-title":"J. Cell Biol"},{"key":"pcbi.1010765.ref035","doi-asserted-by":"crossref","first-page":"e16159","DOI":"10.7554\/eLife.16159","article-title":"Human kinetochores are swivel joints that mediate microtubule attachments","volume":"5","author":"CA Smith","year":"2016","journal-title":"eLife"},{"issue":"430","key":"pcbi.1010765.ref036","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1080\/01621459.1995.10476572","article-title":"Bayes Factors","volume":"90","author":"RE Kass","year":"1995","journal-title":"Journal of the American Statistical Association"},{"issue":"6","key":"pcbi.1010765.ref037","doi-asserted-by":"crossref","first-page":"1623","DOI":"10.1083\/jcb.201607096","article-title":"Congressing kinetochores progressively load Ska complexes to prevent force-dependent detachment","volume":"216","author":"P Auckland","year":"2017","journal-title":"J. Cell Biol"},{"key":"pcbi.1010765.ref038","doi-asserted-by":"crossref","DOI":"10.1007\/978-981-13-2971-5","volume-title":"Monte Carlo Methods","author":"A Barbu","year":"2020"},{"issue":"49","key":"pcbi.1010765.ref039","doi-asserted-by":"crossref","first-page":"17408","DOI":"10.1073\/pnas.1408184111","article-title":"A general construction for parallelizing Metropolis-Hastings algorithms","volume":"111","author":"B Calderhead","year":"2014","journal-title":"PNAS"},{"issue":"8","key":"pcbi.1010765.ref040","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1016\/j.tcb.2020.04.006","article-title":"Crowning the Kinetochore: The Fibrous Corona in Chromosome Segregation","volume":"30","author":"GJPL Kops","year":"2020","journal-title":"Trends in Cell Biology"},{"key":"pcbi.1010765.ref041","doi-asserted-by":"crossref","first-page":"e21007","DOI":"10.7554\/eLife.21007","article-title":"Molecular basis of outer kinetochore assembly on CENP-T","volume":"5","author":"PJ Huis in\u2019t Veld","year":"2016","journal-title":"eLife"},{"key":"pcbi.1010765.ref042","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1146\/annurev-physchem-032511-143716","article-title":"DEER Distance Measurements on Proteins","volume":"63","author":"G Jeschke","year":"2012","journal-title":"Annual Review of Physical Chemistry"},{"key":"pcbi.1010765.ref043","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1146\/annurev-biophys-051013-022846","article-title":"Split Green Fluorescent Proteins: Scope, Limitations and Outlook","volume":"48","author":"MG Romei","year":"2019","journal-title":"Annual Review of Biophysics"}],"updated-by":[{"DOI":"10.1371\/journal.pcbi.1010765","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2023,1,9]],"date-time":"2023-01-09T00:00:00Z","timestamp":1673222400000}}],"container-title":["PLOS Computational Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pcbi.1010765","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,9]],"date-time":"2023-01-09T14:05:41Z","timestamp":1673273141000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pcbi.1010765"}},"subtitle":[],"editor":[{"given":"Melissa L.","family":"Kemp","sequence":"first","affiliation":[]}],"short-title":[],"issued":{"date-parts":[[2022,12,27]]},"references-count":43,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2022,12,27]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pcbi.1010765","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2021.08.22.457266","asserted-by":"object"}]},"ISSN":["1553-7358"],"issn-type":[{"value":"1553-7358","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,27]]}}}