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Whole Ovary Immunofluorescence, Clearing, and Multiphoton Microscopy for Quantitative 3D Analysis of the Developing Ovarian Reserve in Mouse
Bu içeriği görüntülemek için JoVE aboneliği gereklidir. Oturum açın veya ücretsiz deneme sürümünü başlatın.
Whole Ovary Immunofluorescence, Clearing, and Multiphoton Microscopy for Quantitative 3D Analysis of the Developing Ovarian Reserve in Mouse
Bölümler
- 00:04Introduction
- 01:00Perfusion, Dissection, and Fixation of Pubertal Ovaries
- 02:34Whole-Mount Ovary Immunostaining
- 03:59Clearing of Immunostained Whole-Mount Ovaries
- 04:54Sample Setup and Imaging with a Multiphoton Microscope
- 06:54Oocyte Quantification
- 09:28Results: Quantitative and Qualitative Analysis of the Ovaries Using Immunostaining, Multiphoton Microscopy, and 3D Visualization
- 11:51Conclusion
Here, we present an optimized protocol for imaging entire ovaries for quantitative and qualitative analyses using whole-mount immunostaining, multiphoton microscopy, and 3D visualization and analysis. This protocol accommodates high-throughput, reliable, and repeatable processing that is applicable for toxicology, clinical diagnostics, and genomic assays of ovarian function.
