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

Gregory D. Scott, Emily D. Blum, Allison D. Fryer, David B. Jacoby

Research output: Contribution to journalArticle

22 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)43-55
Number of pages13
JournalAmerican journal of respiratory cell and molecular biology
Volume51
Issue number1
DOIs
StatePublished - Jul 2014

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Keywords

  • Clearing
  • Modeling
  • Morphometry
  • Nerve
  • Visceral pleura

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

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