LiverClear: A versatile protocol for mouse liver tissue clearing

• Published in: STAR protocols Vol. 3; no. 1; p. 101178
• Main Authors: Molina, Laura M., Krutsenko, Yekaterina, Jenkins, Nathaniel E.C., Smith, Megan C., Tao, Junyan, Wheeler, Travis B., Watkins, Simon C., Watson, Alan M., Monga, Satdarshan P.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18-03-2022; Elsevier
• Subjects: Animals; Developmental biology; Liver - diagnostic imaging; Mice; Microscopy; Protocol; Staining and Labeling; Microscopy; Developmental biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101178

Although there are numerous tissue clearing protocols, most are inadequate for clearing liver tissue. Here we present a flexible protocol for mouse liver tissue; we combine strategies from several previously published protocols for delipidation, decolorization, staining, and refractive index matching. LiverClear is sufficiently versatile to allow clearing of healthy and diseased mouse liver followed by immunofluorescence staining and imaging to visualize intact 3D structures such as bile ducts and hepatocyte canaliculi. We also adapted this protocol for clearing human livers.
For complete details on the use and execution of this protocol, please refer to Molina et al. (2021).
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•Step-by-step optimized protocol for tissue clearing of healthy or diseased liver•Visualization of livers cells in their 3-dimensional tissue organization•Adaptation of tissue clearing protocol for human livers•Immunostaining and ribbon-scanning confocal microscopy on cleared liver tissue
Although there are numerous tissue clearing protocols, most are inadequate for clearing liver tissue. Here we present a flexible protocol for mouse liver tissue; we combine strategies from several previously published protocols for delipidation, decolorization, staining, and refractive index matching. LiverClear is sufficiently versatile to allow clearing of healthy and diseased mouse liver followed by immunofluorescence staining and imaging to visualize intact 3D structures such as bile ducts and hepatocyte canaliculi. We also adapted this protocol for clearing human livers.