Multi-immersion open-top light-sheet microscope for high-throughput imaging of cleared tissues

Recent advances in optical clearing and light-sheet microscopy have provided unprecedented access to structural and molecular information from intact tissues. However, current light-sheet microscopes have imposed constraints on the size, shape, number of specimens, and compatibility with various clearing protocols. Here we present a multi-immersion open-top light-sheet microscope that enables simple mounting of multiple specimens processed with a variety of protocols, which will facilitate wider adoption by preclinical researchers and clinical laboratories. One Sentence Summary Glaser et al. describe a multi-immersion open-top light-sheet microscope that enables simple and high-throughput imaging of large numbers of preclinical and clinical specimens prepared with a variety of clearing protocols.

[1]  K. Deisseroth,et al.  Advanced CLARITY for rapid and high-resolution imaging of intact tissues , 2014, Nature Protocols.

[2]  N. Renier,et al.  iDISCO: A Simple, Rapid Method to Immunolabel Large Tissue Samples for Volume Imaging , 2014, Cell.

[3]  K. Deisseroth,et al.  CLARITY for mapping the nervous system , 2013, Nature Methods.

[4]  Stephan Preibisch,et al.  BigStitcher: Reconstructing high-resolution image datasets of cleared and expanded samples , 2018 .

[5]  Su Guo,et al.  Open-top selective plane illumination microscope for conventionally mounted specimens. , 2015, Optics express.

[6]  Andreas S Tolias,et al.  Multimodal profiling of single-cell morphology, electrophysiology, and gene expression using Patch-seq , 2017, Nature Protocols.

[7]  Salil S. Bhate,et al.  Deep Profiling of Mouse Splenic Architecture with CODEX Multiplexed Imaging , 2017, Cell.

[8]  Edward S. Boyden,et al.  Expansion microscopy , 2015, Science.

[9]  N. Plesnila,et al.  Shrinkage-mediated imaging of entire organs and organisms using uDISCO , 2016, Nature Methods.

[10]  Michael Z. Lin,et al.  A Suite of Transgenic Driver and Reporter Mouse Lines with Enhanced Brain-Cell-Type Targeting and Functionality , 2018, Cell.

[11]  Ronald N. Germain,et al.  Multiplex, quantitative cellular analysis in large tissue volumes with clearing-enhanced 3D microscopy (Ce3D) , 2017, Proceedings of the National Academy of Sciences.

[12]  Jonas Ries,et al.  A real-time compression library for microscopy images , 2017, bioRxiv.

[13]  Etsuo A. Susaki,et al.  CUBIC pathology: three-dimensional imaging for pathological diagnosis , 2017, Scientific Reports.

[14]  Frank Bradke,et al.  Three-dimensional imaging of solvent-cleared organs using 3DISCO , 2012, Nature Protocols.

[15]  R. Weiler,et al.  Chemical Clearing and Dehydration of GFP Expressing Mouse Brains , 2012, PloS one.

[16]  R. Yuste,et al.  Light sheet theta microscopy for rapid high-resolution imaging of large biological samples , 2018, BMC Biology.

[17]  Jeff W. Lichtman,et al.  Clarifying Tissue Clearing , 2015, Cell.

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

[19]  Aaron S. Andalman,et al.  Structural and molecular interrogation of intact biological systems , 2013, Nature.

[20]  Nils Norlin,et al.  Inverted light-sheet microscope for imaging mouse pre-implantation development , 2015, Nature Methods.

[21]  Karl Deisseroth,et al.  Whole-tissue biopsy phenotyping of three-dimensional tumours reveals patterns of cancer heterogeneity , 2017, Nature Biomedical Engineering.

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

[23]  Chengbo Yin,et al.  Multidirectional digital scanned light-sheet microscopy enables uniform fluorescence excitation and contrast-enhanced imaging , 2018, Scientific Reports.

[24]  Joachim Hornegger,et al.  Virtual Hematoxylin and Eosin Transillumination Microscopy Using Epi-Fluorescence Imaging , 2016, PloS one.

[25]  Lawrence D. True,et al.  Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens , 2017, Nature Biomedical Engineering.

[26]  Kwanghun Chung,et al.  Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues , 2016, Nature Biotechnology.

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

[28]  Aaron R. Halpern,et al.  Volumetric, Nanoscale Optical Imaging of Mouse and Human Kidney via Expansion Microscopy , 2018, Scientific Reports.

[29]  Kevin W Eliceiri,et al.  NIH Image to ImageJ: 25 years of image analysis , 2012, Nature Methods.

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

[31]  Anna Medyukhina,et al.  Fully Automated Evaluation of Total Glomerular Number and Capillary Tuft Size in Nephritic Kidneys Using Lightsheet Microscopy. , 2017, Journal of the American Society of Nephrology : JASN.

[32]  A. Schierloh,et al.  Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain , 2007, Nature Methods.

[33]  Kwanghun Chung,et al.  Simultaneous protection of tissue physicochemical properties using polyfunctional crosslinkers , 2018, Nature Biotechnology.

[34]  William E. Allen,et al.  Three-dimensional intact-tissue sequencing of single-cell transcriptional states , 2018, Science.

[35]  Joshua C Vaughan,et al.  Expansion microscopy with conventional antibodies and fluorescent proteins , 2016, Nature Methods.

[36]  Dimitri Perrin,et al.  Advanced CUBIC protocols for whole-brain and whole-body clearing and imaging , 2015, Nature Protocols.

[37]  Haruo Kasai,et al.  Chemical Landscape for Tissue Clearing Based on Hydrophilic Reagents. , 2018, Cell reports.