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Updated: Mar 22, 2026
Whole Ovary Immunofluorescence, Clearing, and Multiphoton Microscopy for Quantitative 3D Analysis of the Developing Ovarian Reserve in Mouse
Published on: September 3, 2021
Protocol to study murine ovarian elasticity and composition in situ by integrating quantitative micro-elastography
Anna Jaeschke1, Matt S Hepburn2, Alireza Mowla3
1Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore; Center for Neuromusculoskeletal Restorative Medicine, Hong Kong Science Park, Shatin, NT, Hong Kong SAR, China; School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Ma Liu Shui, NT, Hong Kong SAR, China.
This study introduces a new protocol combining quantitative micro-elastography (QME) with microscopy to analyze ovarian elasticity and composition. The method allows for detailed in situ examination of ovarian tissue structure and molecular makeup.
Area of Science:
- Biomedical Engineering
- Reproductive Biology
- Microscopy Techniques
Background:
- Ovarian tissue analysis requires methods that can assess both mechanical properties and molecular composition.
- Existing techniques may not offer sufficient resolution or in situ capabilities for comprehensive study.
Purpose of the Study:
- To present a novel protocol integrating quantitative micro-elastography (QME) with transmitted light (TL) and immunofluorescence (IF) microscopy.
- To enable in situ investigation of ovarian elasticity and composition in murine models.
Main Methods:
- The protocol involves excising murine ovaries, performing QME scans, and tissue fixation.
- Subsequent steps include TL microscopy for identifying ovarian structures (corpora lutea, follicles) and IF microscopy for cellular and molecular analysis.
- Integration of TL and IF imaging with QME data allows for spatially resolved characterization.
Main Results:
- The combined QME-TL-IF approach provides a comprehensive dataset on ovarian mechanical properties and composition.
- Successful segmentation of functional ovarian components within QME data is demonstrated.
- The protocol facilitates detailed in situ analysis of tissue structure and molecular markers.
Conclusions:
- This integrated protocol offers a powerful tool for studying ovarian biology and disease.
- It enables simultaneous assessment of mechanical properties and molecular composition in situ.
- The method has potential applications in reproductive research and diagnostics.
