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Whole Ovary Immunofluorescence, Clearing, and Multiphoton Microscopy for Quantitative 3D Analysis of the Developing Ovarian Reserve in Mouse
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Chapters
- 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.
