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Skeletal Muscle Tissue Clearing for LacZ and Fluorescent Reporters, and Immunofluorescence Staining

  • Mayank Verma
  • Bhavani SR Murkonda
  • Yoko Asakura
  • Atsushi AsakuraEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1460)

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.

Key words

Skeletal muscle Tissue imaging tdTomato Green fluorescence protein (GFP) Satellite cell Tissue clearing lacZ reporter 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Mayank Verma
    • 1
    • 2
    • 3
    • 4
  • Bhavani SR Murkonda
    • 2
    • 3
    • 4
  • Yoko Asakura
    • 2
    • 3
    • 4
  • Atsushi Asakura
    • 2
    • 3
    • 4
    Email author
  1. 1.Medical Scientist Training ProgramUniversity of Minnesota Medical SchoolMinneapolisUSA
  2. 2.Stem Cell InstituteUniversity of Minnesota Medical SchoolMinneapolisUSA
  3. 3.Paul & Sheila Wellstone Muscular Dystrophy CenterUniversity of Minnesota Medical SchoolMinneapolisUSA
  4. 4.Department of NeurologyUniversity of Minnesota Medical SchoolMinneapolisUSA

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