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.

Tissue clearing for confocal imaging of native and bio-artificial skeletal muscle / Decroix, L; Van Muylder, V.; Desender, L.; Sampaolesi, Maurilio; Thorrez, L.. - In: BIOTECHNIC & HISTOCHEMISTRY. - ISSN 1052-0295. - 90:6(2015), pp. 424-431. [10.3109/10520295.2015.1019564]

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

SAMPAOLESI, MAURILIO;
2015

Abstract

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.
2015
bio-artifi cial; clearing; confocal microscopy; skeletal muscle; tissue engineering; visualization; animals; cells; cultured; green fluorescent proteins; human umbilical vein endothelial cells; humans; imaging; three-dimensional; immunohistochemistry; mice; transgenic; microscopy; confocal; muscle; skeletal; myoblasts; skeletal; recombinant proteins; tissue engineering; bioartificial organs; histology; medical laboratory technology
01 Pubblicazione su rivista::01a Articolo in rivista
Tissue clearing for confocal imaging of native and bio-artificial skeletal muscle / Decroix, L; Van Muylder, V.; Desender, L.; Sampaolesi, Maurilio; Thorrez, L.. - In: BIOTECHNIC & HISTOCHEMISTRY. - ISSN 1052-0295. - 90:6(2015), pp. 424-431. [10.3109/10520295.2015.1019564]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1581889
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