Imaging multicellular specimens with real-time optimized tiling light-sheet selective plane illumination microscopy

Despite the progress made in selective plane illumination microscopy, high-resolution 3D live imaging of multicellular specimens remains challenging. Tiling light-sheet selective plane illumination microscopy (TLS-SPIM) with real-time light-sheet optimization was developed to respond to the challenge. It improves the 3D imaging ability of SPIM in resolving complex structures and optimizes SPIM live imaging performance by using a real-time adjustable tiling light sheet and creating a flexible compromise between spatial and temporal resolution. We demonstrate the 3D live imaging ability of TLS-SPIM by imaging cellular and subcellular behaviours in live C. elegans and zebrafish embryos, and show how TLS-SPIM can facilitate cell biology research in multicellular specimens by studying left-right symmetry breaking behaviour of C. elegans embryos.

[1]  Philipp J. Keller,et al.  Imaging Morphogenesis: Technological Advances and Biological Insights , 2013, Science.

[2]  K. Dholakia,et al.  Light-sheet microscopy using an Airy beam , 2014, Nature Methods.

[3]  Zhirong Bao,et al.  Chiral forces organize left-right patterning in C. elegans by uncoupling midline and anteroposterior axis. , 2010, Developmental cell.

[4]  Philipp J. Keller,et al.  Whole-animal functional and developmental imaging with isotropic spatial resolution , 2015, Nature Methods.

[5]  Gaudenz Danuser,et al.  Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy , 2015, Biophysical journal.

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

[7]  Daniel A. Colón-Ramos,et al.  Inverted selective plane illumination microscopy (iSPIM) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans , 2011, Proceedings of the National Academy of Sciences.

[8]  M. Davidson,et al.  Noninvasive Imaging beyond the Diffraction Limit of 3D Dynamics in Thickly Fluorescent Specimens , 2012, Cell.

[9]  Lars Hufnagel,et al.  Multiview light-sheet microscope for rapid in toto imaging , 2012, Nature Methods.

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

[11]  C. Pohl,et al.  Coupling of rotational cortical flow, asymmetric midbody positioning, and spindle rotation mediates dorsoventral axis formation in C. elegans. , 2014, Developmental cell.

[12]  A. Rohrbach,et al.  Microscopy with self-reconstructing beams , 2010 .

[13]  Clemens F. Kaminski,et al.  Fast imaging of live organisms with sculpted light sheets , 2015, Scientific Reports.

[14]  Liang Gao,et al.  Optimization of the excitation light sheet in selective plane illumination microscopy. , 2015, Biomedical optics express.

[15]  Liang Gao,et al.  3D live fluorescence imaging of cellular dynamics using Bessel beam plane illumination microscopy , 2014, Nature Protocols.

[16]  M. Davidson,et al.  Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination , 2011, Nature Methods.

[17]  Erhard Klotz,et al.  Coherent Optical Generation and Inspection of Two-dimensional Periodic Structures , 1977 .

[18]  Liangyi Chen,et al.  Large-field high-resolution two-photon digital scanned light-sheet microscopy , 2014, Cell Research.

[19]  Philipp J. Keller,et al.  Reconstruction of Zebrafish Early Embryonic Development by Scanned Light Sheet Microscopy , 2008, Science.

[20]  Wesley R. Legant,et al.  Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution , 2014, Science.

[21]  Justin Senseney,et al.  Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy , 2013, Nature Biotechnology.

[22]  Philipp J. Keller,et al.  Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy , 2012, Nature Methods.

[23]  Liang Gao,et al.  Extend the field of view of selective plan illumination microscopy by tiling the excitation light sheet. , 2015, Optics express.

[24]  C. Zhou,et al.  Numerical study of Dammann array illuminators. , 1995, Applied optics.

[25]  S. Grill,et al.  Active torque generation by the actomyosin cell cortex drives left–right symmetry breaking , 2014, eLife.

[26]  Christian Pohl,et al.  Cytoskeletal Symmetry Breaking and Chirality: From Reconstituted Systems to Animal Development , 2015, Symmetry.