Dynamic and non-contact 3D sample rotation for microscopy

Precise sample orientation is crucial for microscopy but is often performed with macroscopic tools and low accuracy. In vivo imaging of growing and developing samples even requires dynamic adaptation of the sample orientation to continuously achieve optimal imaging. Here, we present a method for freely positioning a sample in 3D by introducing magnetic beads and applying a magnetic field. We demonstrate magnetic orientation of fixed mouse embryos and artemia, and live zebrafish embryos and larvae on an epi-fluorescence microscope and on a light-sheet system for optimal imaging.Sample orientation is crucial to ensure optimal image quality in light microscopy. Here the authors enable multi-axis orientation of fixed mouse embryos and shrimp, and live zebrafish embryos and larvae by introducing magnetic beads and rotating the sample with a magnetic field in a microscope.

[1]  D. Stainier,et al.  Cellular and molecular analyses of vascular tube and lumen formation in zebrafish , 2005, Development.

[2]  Catarina Brito,et al.  SPIM-fluid: open source light-sheet based platform for high-throughput imaging. , 2015, Biomedical optics express.

[3]  Keir C. Neuman,et al.  Single-Molecule Micromanipulation Techniques , 2007 .

[4]  J. Huisken,et al.  A guide to light-sheet fluorescence microscopy for multiscale imaging , 2017, Nature Methods.

[5]  Christian Dietrich,et al.  The optical cell rotator. , 2008, Optics express.

[6]  Thomas Brox,et al.  ViBE-Z: a framework for 3D virtual colocalization analysis in zebrafish larval brains , 2012, Nature Methods.

[7]  Ralf Dahm,et al.  Zebrafish: A Practical Approach. Edited by C. NÜSSLEIN-VOLHARD and R. DAHM. Oxford University Press. 2002. 322 pages. ISBN 0 19 963808 X. Price £40.00 (paperback). ISBN 0 19 963809 8. Price £80.00 (hardback). , 2003 .

[8]  Tobias Pietzsch,et al.  BigDataViewer: visualization and processing for large image data sets , 2015, Nature Methods.

[9]  Stephan Saalfeld,et al.  Software for bead-based registration of selective plane illumination microscopy data , 2010, Nature Methods.

[10]  J. Campos-Ortega,et al.  A zebrafish histone variant H2A.F/Z and a transgenic H2A.F/Z:GFP fusion protein for in vivo studies of embryonic development , 2001, Development Genes and Evolution.

[11]  K. Lyons,et al.  Whole-mount skeletal staining. , 2014, Methods in molecular biology.

[12]  V. Cunliffe.  Zebrafish: A Practical Approach. Edited by C. NÜSSLEIN-VOLHARD and R. DAHM. Oxford University Press. 2002. 322 pages. ISBN 0 19 963808 X. Price £40.00 (paperback). ISBN 0 19 963809 8. Price £80.00 (hardback). , 2003 .

[13]  Oliver Otto,et al.  Dynamic operation of optical fibres beyond the single-mode regime facilitates the orientation of biological cells , 2014, Nature Communications.

[14]  Fei Wang,et al.  High-throughput mapping of brain-wide activity in awake and drug-responsive vertebrates. , 2015, Lab on a chip.

[15]  Citlali Pérez Campos,et al.  High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics , 2013, Nature Communications.

[16]  F. Del Bene,et al.  Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy , 2004, Science.