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3D imaging of whole cleared endocrine glands and human tissues. (A) Light sheet image of Dupuytren’s disease cord tissue immunostained with SOX9 and Endoglin (left) and α-SMA (right). Nuclei: DAPI. (B) 3D imaging of cleared human liver with α-SMA immunostaining. (C) 3D confocal images of brain tissue before and after tissue clearing, obtained from Cdh5(PAC)-CreERT2; ROSA26-td-Tomato mice and Flk1 GFP reporter mice. (D) 3D images of cleared mouse ovary and adrenal gland stained with CD102 from Cdh5(PAC)-CreERT2 ; ROSA26-td-Tomato mutant mice acquired by confocal microscopy. Nuclei: DAPI. (E) 3D images of cleared ovary and adrenal gland immunostained with BCAM from Flk1 GFP reporter mice. Nuclei: DAPI. (F) 3D images of mouse brain tissue at 0, 3, 7, and 14 d after tissue clearing. Quantification of normalized mean fluorescence ( i d/0 d) ( n = 5). Scale bars: 400 μm (A and B); 50 μm (C and F); 150 μm for tile scan images and 50 μm for high-magnification insets (D and E). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests. ns, not significant.

Journal: Computational and Structural Biotechnology Journal

Article Title: Rapid 3D Immunolabeling and Light Sheet Microscopy for Quantitative Analysis of Intact Tissues

doi: 10.34133/csbj.0121

Figure Lengend Snippet: 3D imaging of whole cleared endocrine glands and human tissues. (A) Light sheet image of Dupuytren’s disease cord tissue immunostained with SOX9 and Endoglin (left) and α-SMA (right). Nuclei: DAPI. (B) 3D imaging of cleared human liver with α-SMA immunostaining. (C) 3D confocal images of brain tissue before and after tissue clearing, obtained from Cdh5(PAC)-CreERT2; ROSA26-td-Tomato mice and Flk1 GFP reporter mice. (D) 3D images of cleared mouse ovary and adrenal gland stained with CD102 from Cdh5(PAC)-CreERT2 ; ROSA26-td-Tomato mutant mice acquired by confocal microscopy. Nuclei: DAPI. (E) 3D images of cleared ovary and adrenal gland immunostained with BCAM from Flk1 GFP reporter mice. Nuclei: DAPI. (F) 3D images of mouse brain tissue at 0, 3, 7, and 14 d after tissue clearing. Quantification of normalized mean fluorescence ( i d/0 d) ( n = 5). Scale bars: 400 μm (A and B); 50 μm (C and F); 150 μm for tile scan images and 50 μm for high-magnification insets (D and E). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests. ns, not significant.

Article Snippet: For experiments involving endogenous fluorescence, Kdr tm2.1Jrt /J mice (stock no.: 017006, Jackson Laboratory) ( ) and ROSA26 td-Tomato reporter mice (stock no.: 007914, Jackson Laboratory) ( ) were used.

Techniques: 3D Imaging, Immunostaining, Staining, Mutagenesis, Confocal Microscopy, Fluorescence, Derivative Assay