12/27/2022 0 Comments Twistit motors and transmission![]() Neuroinformatics of the Allen mouse brain connectivity atlas. Brain tissue scanner enables brain microstructure surveys. Serial two-photon tomography for automated ex vivo mouse brain imaging. Functional targeted brain endoskeletonization. High-resolution imaging of entire organs by 3-dimensional imaging of solvent cleared organs (3DISCO). Light microscopy mapping of connections in the intact brain. Idisco: a simple, rapid method to immunolabel large tissue samples for volume imaging. Optical clearing of fixed brain samples using SeeDB. Whole-body imaging with single-cell resolution by tissue decolorization. A rapid optical clearing protocol using 2,2′-thiodiethanol for microscopic observation of fixed mouse brain. Functional imaging in bulk tissue specimens using optical emission tomography: fluorescence preservation during optical clearing. Sakhalkar, H.S., Dewhirst, M., Oliver, T., Cao, Y. Single-cell phenotyping within transparent intact tissue through whole-body clearing. Scalable and DiI-compatible optical clearance of the mammalian brain. Neuronal calcium-binding proteins 1/2 localize to dorsal root ganglia and excitatory spinal neurons and are regulated by nerve injury. Advanced clarity for rapid and high-resolution imaging of intact tissues. Three-dimensional imaging of solvent-cleared organs using 3DISCO. Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Chemical clearing and dehydration of GFP-expressing mouse brains. Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis. Clear T: a detergent-and solvent-free clearing method for neuronal and non-neuronal tissue. Sca le: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain. Structural and molecular interrogation of intact biological systems. SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction. ![]() Animal models of human disease: zebrafish swim into view. Transparent adult zebrafish as a tool for in vivo transplantation analysis. Emergence of reproducible spatiotemporal activity during motor learning. Optimization of a GCaMP calcium indicator for neural activity imaging. ![]() Ultrasensitive fluorescent proteins for imaging neuronal activity. Together, these protocols and solutions enable phenotyping of subcellular components and tracing cellular connectivity in intact biological networks.Ĭhen, T.W. These methods are adaptable to difficult-to-image tissues, such as bone (PACT-deCAL), and to magnified single-cell visualization (ePACT). Furthermore, the macromolecule permeability of PACT- and PARS-processed tissue hybrids supports the diffusion of immunolabels throughout intact tissue, whereas RIMS (refractive index matching solution) grants high-resolution imaging at depth by further reducing light scattering in cleared and uncleared samples alike. PACT (passive CLARITY technique) and PARS (perfusion-assisted agent release in situ) use tissue-hydrogel hybrids to stabilize tissue biomolecules during selective lipid extraction, resulting in enhanced clearing efficiency and sample integrity. To facilitate fine-scale phenotyping of whole specimens, we describe here a set of tissue fixation-embedding, detergent-clearing and staining protocols that can be used to transform excised organs and whole organisms into optically transparent samples within 1–2 weeks without compromising their cellular architecture or endogenous fluorescence. ![]()
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