Advanced CLARITY for rapid and high-resolution imaging of intact tissues
TLDR
Protocols spanning multiple dimensions of the CLARITY workflow are described, ranging from simple, reliable and efficient lipid removal without electrophoretic instrumentation to optimized objectives and integration with light-sheet optics (CLARITY-optimized light- sheet microscopy (COLM)) for accelerating data collection from clarified samples by several orders of magnitude while maintaining or increasing quality and resolution.Abstract:
CLARITY enables the chemical transformation of intact biological tissues into a hydrogel–tissue hybrid. The hybrid samples can be interrogated using light and macromolecular labels, whilst retaining fine structure and native biological molecules.read more
Citations
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iDISCO: A Simple, Rapid Method to Immunolabel Large Tissue Samples for Volume Imaging
TL;DR: iDISCO enables facile volume imaging of immunolabeled structures in complex tissues and reveals unexpected variability in number of apoptotic neurons within individual sensory ganglia despite tight control of total number in all ganglia.
Journal ArticleDOI
Clarifying Tissue Clearing.
TL;DR: The physical basis for light scatter in tissue is introduced, the mechanisms underlying various clearing techniques are described, and several of the major advances in light microscopy for imaging cleared tissue are discussed.
Journal ArticleDOI
Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing
Bin Yang,Jennifer B. Treweek,Rajan P. Kulkarni,Rajan P. Kulkarni,Benjamin E. Deverman,Chun-Kan Chen,Eric Lubeck,Sheel Shah,Long Cai,Viviana Gradinaru +9 more
TL;DR: Techniques for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically transparent, thereby exposing their cellular structure with intact connectivity are presented.
Journal ArticleDOI
Intact-Brain Analyses Reveal Distinct Information Carried by SNc Dopamine Subcircuits.
Talia N. Lerner,Carrie Shilyansky,Thomas J. Davidson,Kathryn E. Evans,Kevin T. Beier,Kelly A. Zalocusky,Ailey K. Crow,Robert C. Malenka,Liqun Luo,Raju Tomer,Karl Deisseroth +10 more
TL;DR: Two parallel nigrostriatal dopamine neuron subpopulations differing in biophysical properties, input wiring, output wiring to dorsomedial striatum versus dorsolateral striatum (DLS), and natural activity patterns during free behavior are identified.
Journal ArticleDOI
Advanced CUBIC protocols for whole-brain and whole-body clearing and imaging.
Etsuo A. Susaki,Etsuo A. Susaki,Etsuo A. Susaki,Kazuki Tainaka,Kazuki Tainaka,Kazuki Tainaka,Dimitri Perrin,Hiroko Yukinaga,Akihiro Kuno,Hiroki R. Ueda,Hiroki R. Ueda,Hiroki R. Ueda +11 more
TL;DR: A protocol for advanced CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis) is described in this paper, which enables simple and efficient organ clearing, rapid imaging by light-sheet microscopy and quantitative imaging analysis of multiple samples.
References
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Deep tissue two-photon microscopy
Fritjof Helmchen,Winfried Denk +1 more
TL;DR: Fundamental concepts of nonlinear microscopy are reviewed and conditions relevant for achieving large imaging depths in intact tissue are discussed.
Journal ArticleDOI
Optical sectioning deep inside live embryos by selective plane illumination microscopy
TL;DR: In this article, a selective plane illumination microscopy (SPIM) was developed to generate multidimensional images of samples up to a few millimeters in size, which can be applied to visualize the embryogenesis of the relatively opaque Drosophila melanogaster in vivo.
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Structural and molecular interrogation of intact biological systems
Kwanghun Chung,Jenelle Wallace,Sung-Yon Kim,Sandhiya Kalyanasundaram,Aaron S. Andalman,Thomas J. Davidson,Julie J. Mirzabekov,Kelly A. Zalocusky,Joanna Mattis,Aleksandra K. Denisin,Sally Pak,Hannah L. Bernstein,Charu Ramakrishnan,Logan Grosenick,Viviana Gradinaru,Karl Deisseroth +15 more
TL;DR: It is shown that CLARITY enables fine structural analysis of clinical samples, including non-sectioned human tissue from a neuropsychiatric-disease setting, establishing a path for the transmutation of human tissue into a stable, intact and accessible form suitable for probing structural and molecular underpinnings of physiological function and disease.
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Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy.
TL;DR: This work developed digital scanned laser light sheet fluorescence microscopy and recorded nuclei localization and movement in entire wild-type and mutant zebrafish embryos over the first 24 hours of development to derive a model of germ layer formation and show that the mesendoderm forms from one-third of the embryo's cells in a single event.
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Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain.
Hans-Ulrich Dodt,Ulrich Leischner,Anja Schierloh,Nina Jährling,Nina Jährling,Christoph P. Mauch,Katrin Deininger,Jan M. Deussing,Matthias Eder,Walter Zieglgänsberger,Klaus Becker,Klaus Becker +11 more
TL;DR: This new technique allows optical sectioning of fixed mouse brains with cellular resolution and can be used to detect single GFP-labeled neurons in excised mouse hippocampi and is ideally suited for high-throughput phenotype screening of transgenic mice and thus will benefit the investigation of disease models.