title: Skeletal Muscle Tissue Clearing for LacZ and Fluorescent Reporters, and Immunofluorescence Staining.
abstract: Skeletal muscle is a highly ordered yet complex tissue containing several cell types that interact with each other in order to maintain structure and homeostasis. It is also a highly regenerative tissue that responds to damage in a highly intricate but stereotypic manner, with distinct spatial and temporal kinetics. Proper examination of this process requires one to look at the three-dimensional orientation of the cellular and subcellular components, which can be accomplished through tissue clearing. While there has been a recent surge of protocols to study biology in whole tissue, it has primarily focused on the nervous system. This chapter describes the workflow for whole mount analysis of murine skeletal muscle for LacZ reporters, fluorescent reporters and immunofluorescence staining. Using this technique, we are able to visualize LacZ reporters more effectively in deep tissue samples, and to perform fluorescent imaging with a depth greater than 1700 μm.
authors: Medical Scientist Training Program, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.|Stem Cell Institute, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.|Paul & Sheila Wellstone Muscular Dystrophy Center, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.|Department of Neurology, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.
authors: Mayank
authors: M
authors: Verma
authors: Stem Cell Institute, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.|Paul & Sheila Wellstone Muscular Dystrophy Center, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.|Department of Neurology, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.
authors: Bhavani Sr
authors: BS
authors: Murkonda
authors: Stem Cell Institute, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.|Paul & Sheila Wellstone Muscular Dystrophy Center, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.|Department of Neurology, University of Minnesota Medical School, Minneapolis, MN, 55455, USA.
authors: Yoko
authors: Y
authors: Asakura
authors: Stem Cell Institute, University of Minnesota Medical School, Minneapolis, MN, 55455, USA. asakura@umn.edu.|Paul & Sheila Wellstone Muscular Dystrophy Center, University of Minnesota Medical School, Minneapolis, MN, 55455, USA. asakura@umn.edu.|Department of Neurology, University of Minnesota Medical School, Minneapolis, MN, 55455, USA. asakura@umn.edu.
authors: Atsushi
authors: A
authors: Asakura
mesh_terms: D000818:Animals; D015415:Biomarkers; D005455:Fluorescent Antibody Technique; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D018482:Muscle, Skeletal / Q000378:metabolism*; D013194:Staining and Labeling; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase
keywords: Green fluorescence protein (GFP); Satellite cell; Skeletal muscle; Tissue clearing; Tissue imaging; lacZ reporter; tdTomato