研究目的
To develop and refine a protocol for the immunofluorescent labeling and clarification of the placenta in the nonhuman primate to allow for high-resolution 3D imaging of sufficient quality for advanced quantification methods.
研究成果
The developed protocol allows for the generation of confocal imaged z-stacks with sufficient resolution to pursue reliable and reproducible quantitative analyses of the nonhuman primate placental micro-anatomy. This method provides critical insights into both human and nonhuman primate pregnancies, potentially translating to clinical breakthroughs and improvement in obstetric outcomes.
研究不足
The methodology lacks a signal:noise ratio of sufficient magnitude to allow for quantitative analysis without refinement. The protocol requires optimization for different antigens and may not be directly applicable to other tissues without modification.
1：Experimental Design and Method Selection:
Developed a protocol for the preparation, immunolabeling, and clarification of the nonhuman primate placenta refined for confocal microscopy and subsequent quantification of the microanatomic structures.
2：Sample Selection and Data Sources:
Utilized over 300 samples from 20 nonhuman primate pregnancies.
3：List of Experimental Equipment and Materials:
Included Visikol? HISTO? for tissue clarification, Leica SP5 AOB5 spectral confocal system for imaging, and specific antibodies for immunolabeling.
4：Experimental Procedures and Operational Workflow:
Detailed steps included sample preparation, removal of heme byproducts, antigen retrieval, immunofluorescent labeling, tissue clarification, image acquisition, and 3D rendering.
5：Data Analysis Methods:
Used ImageJ for 3D rendering and quantitative analysis of the placental micro-anatomy.
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