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      Tissue optical clearing, three-dimensional imaging, and computer morphometry in whole mouse lungs and human airways.

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          Abstract

          In whole adult mouse lung, full identification of airway nerves (or other cellular/subcellular objects) has not been possible due to patchy distribution and micron-scale size. Here we describe a method using tissue clearing to acquire the first complete image of three-dimensional (3D) innervation in the lung. We then created a method to pair analysis of nerve (or any other colabeled epitope) images with identification of 3D tissue compartments and airway morphometry by using fluorescent casting and morphometry software (which we designed and are making available as open-source). We then tested our method to quantify a sparse heterogeneous nerve population by examining visceral pleural nerves. Finally, we demonstrate the utility of our method in human tissue to image full thickness innervation in irregular 3D tissue compartments and to quantify sparse objects (intrinsic airway ganglia). Overall, this method can uniquely pair the advantages of whole tissue imaging and cellular/subcellular fluorescence microscopy.

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          Author and article information

          Journal
          Am. J. Respir. Cell Mol. Biol.
          American journal of respiratory cell and molecular biology
          1535-4989
          1044-1549
          Jul 2014
          : 51
          : 1
          Affiliations
          [1 ] Division of Pulmonary and Critical Care, Oregon Health and Sciences University, Portland, Oregon.
          Article
          10.1165/rcmb.2013-0284OC
          4091855
          24471696

          clearing, modeling, morphometry, nerve, visceral pleura

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