Imaging lung regeneration by light sheet microscopy.

Histochemistry and Cell Biology
Isabelle SalwigThomas Braun

Abstract

Optical clearing combined with deep imaging of large biological specimen allows organ-wide visualization of cells in three dimensions (3D) to explore regenerative processes in a spatial context. Here, we investigate the dynamics of airway regeneration following toxin-mediated epithelial injury in cleared whole lung preparations by light sheet microscopy. We use a recently developed knock-in mouse strain labeling bronchiolar Club cells (Scgb1a1-mCherry) to define an optimal clearing procedure that efficiently preserves genetically encoded fluorophores. Dehydration in pH-adjusted tert-butanol followed by clearing in ethyl cinnamate maintained maximum mCherry fluorescence while preventing unfavorable background fluorescence. We apply this technique to depict the course of bronchiolar epithelial renewal from an acute injury phase to early and late recovery stages. 3D reconstructions of whole lungs demonstrate near-complete loss of secretory Club cells throughout the entire respiratory tract 3 days post naphthalene (dpn). Multiple foci of regenerating Club cells emerge at 7 dpn, predominantly at airway bifurcations and in distal terminal bronchioles-anatomical regions assumed to harbor distinct stem/progenitor cells subsets. At 21 d...Continue Reading

References

Oct 6, 1997·American Journal of Respiratory Cell and Molecular Biology·M V FanucchiC G Plopper
Mar 25, 1998·Biophysical Journal·M KneenA S Verkman
Jul 25, 2009·Proceedings of the National Academy of Sciences of the United States of America·Jason R RockBrigid L M Hogan
Aug 8, 2013·The Journal of Clinical Investigation·Christina E BarkauskasBrigid L M Hogan
Jul 18, 2015·Cell·Douglas S Richardson, Jeff W Lichtman
Aug 5, 2016·Journal of the American Society of Nephrology : JASN·Anika KlingbergMatthias Gunzer
Apr 1, 2017·Nature Methods·Rory M Power, Jan Huisken
Jan 23, 2019·Development·Wouter MasselinkElly M Tanaka

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Citations

Feb 12, 2021·Histochemistry and Cell Biology·Christian Mühlfeld, Douglas J Taatjes

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Methods Mentioned

BETA
light sheet microscopy
fluorescence microscopy
transgenic

Software Mentioned

Imaris
LaVision BioTec ImSpector

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