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Application of corrosion cast method for scanning electron microscopic observation of mouse embryo vasculature
S Kondo1, R Suzuki, K Yamazaki
1Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Abstract:
We described a method for making vascular corrosion casts of mouse embryos and applied it to study the vasculature of the whole body mouse embryo at 13.5 days (body length 9-11 mm) to 18.5 days (body length 18-23 mm), as well as that of its respective organs. The Mercox resin was perfused via the umbilical vessels using a special needle made by pulling the hematocrit tube (caliber 100-200 microns). The specimens were treated with 10% NaOH, rinsed, dried, coated with gold and observed under a scanning electron microscope. Visceral organs of a 13.5 day embryo were rather immature, but the liver vasculature was relatively well developed. In a 15.5 day embryo, the whole body vasculature was relatively well developed and the features of respective organs could be identified. In a 18.5 day embryo, almost all organs and extremities were well developed, but sensory organs in some parts showed immature development.
Insights
This study presents a novel method for creating vascular corrosion casts in mouse embryos. This technique reveals detailed whole-body and organ vasculature development from 13.5 to 18.5 days of gestation.
Area of Science:
- Developmental Biology
- Vascular Biology
- Microscopy
Background:
- Understanding embryonic development is crucial for identifying congenital anomalies.
- Detailed visualization of the developing vasculature aids in studying organogenesis and potential vascular defects.
Purpose of the Study:
- To establish and validate a robust method for creating vascular corrosion casts of mouse embryos.
- To characterize the developmental progression of the whole-body and organ-specific vasculature in mouse embryos from 13.5 to 18.5 days of gestation.
Main Methods:
- Perfusing Mercox resin via umbilical vessels into mouse embryos (13.5-18.5 days gestation).
- Utilizing a specialized needle crafted from a hematocrit tube for resin injection.
- Processing specimens with NaOH, followed by rinsing, drying, gold coating, and scanning electron microscopy (SEM).
Main Results:
- The liver vasculature showed significant development by 13.5 days gestation.
- By 15.5 days gestation, whole-body vasculature was well-developed, allowing identification of organ-specific vascular patterns.
- At 18.5 days gestation, most organs and extremities exhibited mature vasculature, though some sensory organs displayed immaturity.
Conclusions:
- The described vascular corrosion casting technique provides high-resolution insights into embryonic vascular development.
- This method is effective for studying the dynamic changes in vasculature during late-stage mouse embryogenesis.
- The findings highlight specific developmental timelines for different organ systems' vascularization.