{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T12:40:45Z","timestamp":1793104845486,"version":"4.5.0"},"reference-count":62,"publisher":"National Academy of Sciences","issue":"46","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2011,11,15]]},"abstract":"<jats:p>\n                    Recently, single-molecule imaging and photocontrol have enabled superresolution optical microscopy of cellular structures beyond Abbe\u2019s diffraction limit, extending the frontier of noninvasive imaging of structures within living cells. However, live-cell superresolution imaging has been challenged by the need to image three-dimensional (3D) structures relative to their biological context, such as the cellular membrane. We have developed a technique, termed superresolution by power-dependent active intermittency and points accumulation for imaging in nanoscale topography (SPRAIPAINT) that combines imaging of intracellular enhanced YFP (eYFP) fusions (SPRAI) with stochastic localization of the cell surface (PAINT) to image two different fluorophores sequentially with only one laser. Simple light-induced blinking of eYFP and collisional flux onto the cell surface by Nile red are used to achieve single-molecule localizations, without any antibody labeling, cell membrane permeabilization, or thiol-oxygen scavenger systems required. Here we demonstrate live-cell 3D superresolution imaging of Crescentin-eYFP, a cytoskeletal fluorescent protein fusion, colocalized with the surface of the bacterium\n                    <jats:italic>Caulobacter crescentus<\/jats:italic>\n                    using a double-helix point spread function microscope. Three-dimensional colocalization of intracellular protein structures and the cell surface with superresolution optical microscopy opens the door for the analysis of protein interactions in living cells with excellent precision (20\u201340\u00a0nm in 3D) over a large field of view (12\u00a0\u00d7\u00a012\u00a0\u03bcm).\n                  <\/jats:p>","DOI":"10.1073\/pnas.1114444108","type":"journal-article","created":{"date-parts":[[2011,10,26]],"date-time":"2011-10-26T21:12:07Z","timestamp":1319663527000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":126,"title":["Three-dimensional superresolution colocalization of intracellular protein superstructures and the cell surface in live\n                    <i>Caulobacter crescentus<\/i>"],"prefix":"10.1073","volume":"108","author":[{"given":"Matthew D.","family":"Lew","sequence":"first","affiliation":[{"name":"Department of Electrical Engineering, Stanford University, Stanford, CA 94305;"},{"name":"Department of Chemistry, Stanford University, Stanford, CA 94305;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven F.","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Stanford University, Stanford, CA 94305;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jerod L.","family":"Ptacin","sequence":"additional","affiliation":[{"name":"Department of Developmental Biology, Stanford University School of Medicine, Beckman Center, Stanford, CA 94305; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marissa K.","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Stanford University, Stanford, CA 94305;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert J.","family":"Twieg","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Kent State University, Kent, OH 44242"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucy","family":"Shapiro","sequence":"additional","affiliation":[{"name":"Department of Developmental Biology, Stanford University School of Medicine, Beckman Center, Stanford, CA 94305; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W. E.","family":"Moerner","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Stanford University, Stanford, CA 94305;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2011,10,26]]},"reference":[{"key":"e_1_1_2_17_8_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2011.02.022"},{"key":"e_1_1_2_17_8_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0609643104"},{"key":"e_1_1_2_17_8_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0900245106"},{"key":"e_1_1_2_17_8_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tetlet.2007.03.026"},{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.e03-07-0522"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1367"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1156947"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0604965103"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1127344"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1529\/biophysj.106.091116"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200802376"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth929"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0609643104"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja1099193"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1258"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1257"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja806590m"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1153529"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0900245106"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1364\/OL.36.000202"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1211"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1004899107"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1364\/OE.17.008264"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/nl903295p"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0813131106"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1583"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1000137"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2083"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2011.02.022"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1146598"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0710517105"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tetlet.2007.03.026"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1084398"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0604503103"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1364\/OE.16.022048"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.3158923"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(03)00935-8"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1795509"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1038\/41048"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.461392"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01081333"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.97.1.151"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1191"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1296"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1298"},{"key":"e_1_3_3_42_2","first-page":"231","article-title":"Biological properties and classification of the caulobacter group","volume":"28","author":"Poindexter JS","year":"1964","unstructured":"JS Poindexter, Biological properties and classification of the caulobacter group. Microbiol Mol Biol Rev 28, 231\u2013295 (1964).","journal-title":"Microbiol Mol Biol Rev"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro864"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.genet.41.110306.130346"},{"key":"e_1_3_3_45_2","doi-asserted-by":"publisher","DOI":"10.1021\/nl103472b"},{"key":"e_1_3_3_46_2","doi-asserted-by":"publisher","DOI":"10.1021\/jp049684d"},{"key":"e_1_3_3_47_2","doi-asserted-by":"publisher","DOI":"10.1021\/ar0401294"},{"key":"e_1_3_3_48_2","doi-asserted-by":"publisher","DOI":"10.1021\/jp907080r"},{"key":"e_1_3_3_49_2","doi-asserted-by":"publisher","DOI":"10.1021\/jp0574145"},{"key":"e_1_3_3_50_2","doi-asserted-by":"publisher","DOI":"10.1021\/jp0534232"},{"key":"e_1_3_3_51_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.3499652"},{"key":"e_1_3_3_52_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1274"},{"key":"e_1_3_3_53_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkm818"},{"key":"e_1_3_3_54_2","doi-asserted-by":"publisher","DOI":"10.1117\/12.842608"},{"key":"e_1_3_3_55_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2818.1993.tb03315.x"},{"key":"e_1_3_3_56_2","doi-asserted-by":"publisher","DOI":"10.1364\/OL.30.001318"},{"key":"e_1_3_3_57_2","doi-asserted-by":"publisher","DOI":"10.1364\/AO.48.001886"},{"key":"e_1_3_3_58_2","doi-asserted-by":"publisher","DOI":"10.1021\/ie071349z"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.1114444108","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T20:41:55Z","timestamp":1649796115000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.1114444108"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,10,26]]},"references-count":62,"journal-issue":{"issue":"46","published-print":{"date-parts":[[2011,11,15]]}},"alternative-id":["10.1073\/pnas.1114444108"],"URL":"https:\/\/doi.org\/10.1073\/pnas.1114444108","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,10,26]]},"assertion":[{"value":"2011-10-26","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}