Tissue clearing for confocal imaging of native and bio-artificial skeletal muscle

Tissue clearing for confocal imaging of native and bio-artificial skeletal muscle

Novel clearing techniques have revolutionized three-dimensional confocal imaging of the brain without the need for physical tissue sectioning. We evaluated three clearing methods, ScaleA2, Clear(T2), and 3DISCO for visualizing native and tissue engineered muscle by confocal microscopy. We found that...

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Bibliographic Details
Published in:Biotechnic & histochemistry Vol. 90; no. 6; p. 424
Main Authors: Decroix, L, Van Muylder, V, Desender, L, Sampaolesi, M, Thorrez, L
Format: Journal Article
Language:English
Published: England 01-01-2015
Subjects:
Animals
Bioartificial Organs
Cells, Cultured
Green Fluorescent Proteins - metabolism
Human Umbilical Vein Endothelial Cells
Humans
Imaging, Three-Dimensional
Immunohistochemistry
Mice
Mice, Transgenic
Microscopy, Confocal - methods
Muscle, Skeletal - anatomy & histology
Muscle, Skeletal - metabolism
Myoblasts, Skeletal - cytology
Myoblasts, Skeletal - metabolism
Recombinant Proteins - metabolism
Tissue Engineering
bio-artificial
tissue engineering
visualization
skeletal muscle
confocal microscopy
clearing
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Summary:Novel clearing techniques have revolutionized three-dimensional confocal imaging of the brain without the need for physical tissue sectioning. We evaluated three clearing methods, ScaleA2, Clear(T2), and 3DISCO for visualizing native and tissue engineered muscle by confocal microscopy. We found that Clear(T2) treatment improved the depth of visualization of immunohistochemical staining slightly, but did not improve depth of visualization of endogenous green fluorescent protein (GFP). ScaleA2 preserved endogenous GFP signal better and permitted significantly deeper GFP imaging, but it was incompatible with tropomyosin immunohistochemical staining. 3DISCO treatment preserved both endogenous GFP and immunohistochemical staining, and permitted significantly deeper imaging. Clearing time for the 3DISCO procedure is short compared to ScaleA2 and Clear(T2). We suggest that 3DISCO is the preferable clearing method for native and tissue engineered skeletal muscle tissue.
ISSN:1473-7760
DOI:10.3109/10520295.2015.1019564