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Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and <t>Prox1</t> immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).
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Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and <t>Prox1</t> immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).
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Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and <t>Prox1</t> immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).
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Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and <t>Prox1</t> immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).
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Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and <t>Prox1</t> immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).
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Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and <t>Prox1</t> immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).
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Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and Prox1 immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).

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: Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and Prox1 immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).

Article Snippet: Quantification of Prox1 + cell numbers was performed using Imaris software (version 9.2.1) based on single-cell resolution 3D images.

Techniques: Imaging, Immunostaining, Expressing, Derivative Assay