RT Journal Article
T1 A Customized Light Sheet Microscope to Measure Spatio-Temporal Protein Dynamics in Small Model Organisms
A1 Ripoll Lorenzo, Jorge
A1 Rieckher, Matthias
A1 Kyparissidis-Kokkinidis, Ilias
A1 Zacharopoulos, Athanasios
A1 Kourmoulakis, Georgios
A1 Zacharakis, Giannis
A1 Tavernarakis, Nektarios
AB 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.
SN 1932-6203
YR 2015
FD 2015-05-22
LK https://hdl.handle.net/10016/20814
UL https://hdl.handle.net/10016/20814
LA eng
NO MR, GZ and AZ acknowledge funding from the Projects “Skin-DOCTor” Grant No. 1778 and “Neureka!” Grant No. LSF7-341 implemented underthe "ARISTEIA" and "Supporting Postdoctoral Researchers" Actions respectively, of the "OPERATIONAL PROGRAMME EDUCATION ANDLIFELONG LEARNING", (http://www.espa.gr/en/pages/staticOPEducationandLifelongLearning.aspx),which is co-funded by the European Social Fund andNational 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), andSpanish 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 themanuscript.
DS e-Archivo
RD 2 may. 2024