{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,3]],"date-time":"2026-02-03T19:14:54Z","timestamp":1770146094440,"version":"3.49.0"},"reference-count":36,"publisher":"Rockefeller University Press","issue":"2","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2016,1,18]]},"abstract":"<jats:p>The understanding of cytoskeleton dynamics has benefited from the capacity to generate fluorescent fiducial marks on cytoskeleton components. Here we show that light-induced imprinting of three-dimensional (3D) fluorescent speckles significantly improves speckle signal and contrast relative to classic (random) fluorescent speckle microscopy. We predict theoretically that speckle imprinting using photobleaching is optimal when the laser energy and fluorophore responsivity are related by the golden ratio. This relation, which we confirm experimentally, translates into a 40% remaining signal after speckle imprinting and provides a rule of thumb in selecting the laser power required to optimally prepare the sample for imaging. This inducible speckle imaging (ISI) technique allows 3D speckle microscopy to be performed in readily available libraries of cell lines or primary tissues expressing fluorescent proteins and does not preclude conventional imaging before speckle imaging. As a proof of concept, we use ISI to measure metaphase spindle microtubule poleward flux in primary cells and explore a scaling relation connecting microtubule flux to metaphase duration.<\/jats:p>","DOI":"10.1083\/jcb.201506128","type":"journal-article","created":{"date-parts":[[2016,2,18]],"date-time":"2016-02-18T23:10:13Z","timestamp":1455837013000},"page":"245-255","update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":6,"title":["Inducible fluorescent speckle microscopy"],"prefix":"10.1083","volume":"212","author":[{"given":"Ant\u00f3nio J.","family":"Pereira","sequence":"first","affiliation":[{"name":"Chromosome Instability and Dynamics Laboratory, Instituto de Biologia Molecular e Celular, Universidade do Porto, 4200-135 Porto, Portugal 1"},{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade \u2013 i3S, Universidade do Porto, 4200-135 Porto, Portugal 2"}]},{"given":"Paulo","family":"Aguiar","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade \u2013 i3S, Universidade do Porto, 4200-135 Porto, Portugal 2"},{"name":"Instituto de Engenharia Biom\u00e9dica, Universidade do Porto, 4200-135 Porto, Portugal 3"}]},{"given":"Michael","family":"Belsley","sequence":"additional","affiliation":[{"name":"Center of Physics, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal 4"}]},{"given":"Helder","family":"Maiato","sequence":"additional","affiliation":[{"name":"Chromosome Instability and Dynamics Laboratory, Instituto de Biologia Molecular e Celular, Universidade do Porto, 4200-135 Porto, Portugal 1"},{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade \u2013 i3S, Universidade do Porto, 4200-135 Porto, Portugal 2"},{"name":"Cell Division Unit, Department of Experimental Biology, Faculty of Medicine, Universidade do Porto, 4200-319 Porto, Portugal 5"}]}],"member":"291","published-online":{"date-parts":[[2016,1,18]]},"reference":[{"key":"2023072309454660100_bib1","doi-asserted-by":"publisher","first-page":"1055","DOI":"10.1016\/S0006-3495(76)85755-4","article-title":"Mobility measurement by analysis of fluorescence photobleaching recovery kinetics","volume":"16","author":"Axelrod","year":"1976","journal-title":"Biophys. 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