A Customized Light Sheet Microscope to Measure Spatio-Temporal Protein Dynamics in Small Model Organisms

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dc.contributor.author Ripoll Lorenzo, Jorge
dc.contributor.author Rieckher, Matthias
dc.contributor.author Kyparissidis-Kokkinidis, Ilias
dc.contributor.author Zacharopoulos, Athanasios
dc.contributor.author Kourmoulakis, Georgios
dc.contributor.author Zacharakis, Giannis
dc.contributor.author Tavernarakis, Nektarios
dc.date.accessioned 2015-05-26T07:33:17Z
dc.date.available 2015-05-26T07:33:17Z
dc.date.issued 2015-05-22
dc.identifier.bibliographicCitation Rieckher M, Kyparissidis-Kokkinidis I, Zacharopoulos A, Kourmoulakis G, Tavernarakis N, Ripoll J, et al. (2015) A Customized Light Sheet Microscope to Measure Spatio-Temporal Protein Dynamics in Small Model Organisms. PLoS ONE 10(5): e0127869. doi:10.1371/journal.pone.0127869
dc.identifier.issn 1932-6203
dc.identifier.uri http://hdl.handle.net/10016/20814
dc.description.abstract We describe a customizable and cost-effective light sheet microscopy (LSM) platform for rapid three-dimensional imaging of protein dynamics in small model organisms. The system is designed for high acquisition speeds and enables extended time-lapse in vivo experiments when using fluorescently labeled specimens. We demonstrate the capability of the setup to monitor gene expression and protein localization during ageing and upon starvation stress in longitudinal studies in individual or small groups of adult Caenorhabditis elegans nematodes. The system is equipped to readily perform fluorescence recovery after photobleaching (FRAP), which allows monitoring protein recovery and distribution under low photobleaching conditions. Our imaging platform is designed to easily switch between light sheet microscopy and optical projection tomography (OPT) modalities. The setup permits monitoring of spatio-temporal expression and localization of ageing biomarkers of subcellular size and can be conveniently adapted to image a wide range of small model organisms and tissue samples.
dc.description.sponsorship MR, GZ and AZ acknowledge funding from the Projects “Skin-DOCTor” Grant No. 1778 and “Neureka!” Grant No. LSF7-341 implemented under the "ARISTEIA" and "Supporting Postdoctoral Researchers" Actions respectively, of the "OPERATIONAL PROGRAMME EDUCATION AND LIFELONG LEARNING", (http://www.espa.gr/en/pages/staticOPEducationandLifelongLearning.aspx), which is co-funded by the European Social Fund and National Resources and from the EU Marie Curie Initial Training Network “OILTEBIA”, Grant No. PITNGA-2012-317526 (http://ec.europa.eu/research/mariecurieactions/, http://gdo.uc3m.es/oiltebia/index. php?option = com_content&view = frontpage). JR acknowledges support from EC FP7 CIG grant HIGHTHROUGHPUT TOMO PICIG12-GA-2012-333632, (http://cordis.europa.eu/projects/333632), and Spanish MINECO grant MESO-IMAGING FIS2013-41802-R, (https://sede.micinn.gob.es/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
dc.format.extent 15
dc.format.mimetype application/pdf
dc.language.iso eng
dc.rights Atribución 3.0 España
dc.rights.uri http://creativecommons.org/licenses/by/3.0/es/
dc.title A Customized Light Sheet Microscope to Measure Spatio-Temporal Protein Dynamics in Small Model Organisms
dc.type article
dc.relation.publisherversion http://dx.doi.org/10.1371/journal.pone.0127869
dc.subject.eciencia Medicina
dc.identifier.doi 10.1371/journal.pone.0127869
dc.rights.accessRights openAccess
dc.relation.projectID info:eu-repo/grantAgreement/EC/FP7/333632
dc.relation.projectID info:eu-repo/grantAgreement/EC/FP7/317526
dc.relation.projectID Gobierno de España. FIS2013-41802-R
dc.type.version publishedVersion
dc.identifier.publicationfirstpage 1
dc.identifier.publicationissue 5
dc.identifier.publicationlastpage 15
dc.identifier.publicationtitle PloS one
dc.identifier.publicationvolume 10
dc.identifier.uxxi AR/0000016872
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