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Structural stabilization of tissue for embryo phenotyping using micro-CT with iodine staining.
Michael D Wong1, Shoshana Spring2, R Mark Henkelman1
1Mouse Imaging Centre, Hospital for Sick Children, Toronto, Ontario, Canada ; Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.
Plos One
|January 4, 2014
Summary
A new hydrogel stabilization method improves mouse embryo imaging by combining it with iodine contrast. This technique reduces tissue distortion and anatomical variation, enhancing the detection of genetic differences in knockout mice.
Area of Science:
- Developmental Biology
- Genomics
- Medical Imaging
Background:
- The International Mouse Phenotyping Consortium aims to understand gene function through large-scale knockout mouse studies.
- Embryonic lethality affects 30% of knockout mouse lines, necessitating advanced phenotyping techniques for late gestation embryos.
- Iodine-enhanced micro-computed tomography (micro-CT) offers superior soft-tissue contrast but causes dehydration and distortion.
Purpose of the Study:
- To develop an improved imaging protocol for mouse embryos that minimizes tissue distortion while maintaining contrast enhancement.
- To reduce inter-sample anatomical variation in late gestation mouse embryos for more sensitive phenotyping.
Main Methods:
- A novel protocol combining a hydrogel mesh with iodinated contrast solutions for micro-CT imaging.
- Formaldehyde fixation to cross-link tissue and hydrogel monomers to biomolecules, stabilizing tissue structure.
- Image analysis using registration methods to quantify anatomical variation.
Main Results:
- The hydrogel stabilization protocol preserves embryo conformation, minimizing distortion caused by iodine solutions.
- Achieved a 20% and 50% reduction in inter-sample variation for normalized brain and lung volumes, respectively.
- Demonstrated a 20% reduction in overall embryo anatomical variation, increasing the sensitivity of automated analysis.
Conclusions:
- Hydrogel stabilization is an effective method to mitigate iodine-induced tissue deformation in mouse embryos.
- This technique enhances the quality and consistency of mouse embryo imaging, improving the ability to detect anatomical differences.
- The protocol supports the International Mouse Phenotyping Consortium's goal of detailed gene function analysis through improved embryo phenotyping.
