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Diffusion microscopic MRI of the mouse embryo: Protocol and practical implementation in the splotch mouse model
Francesca C Norris1, Bernard M Siow, Jon O Cleary
1UCL Centre for Advanced Biomedical Imaging, Division of Medicine, University College London, London, United Kingdom; Centre for Mathematics and Physics in the Life Sciences and EXperimental Biology (CoMPLEX), University College London, London, United Kingdom.
Purpose:
Advanced methodologies for visualizing novel tissue contrast are essential for phenotyping the ever-increasing number of mutant mouse embryos being generated. Although diffusion microscopic MRI (microMRI) has been used to phenotype embryos, widespread routine use is limited by extended scanning times, and there is no established experimental procedure ensuring optimal data acquisition.
Methods:
We developed two protocols for designing experimental procedures for diffusion microMRI of mouse embryos, which take into account the effect of embryo preparation and pulse sequence parameters on resulting data. We applied our protocols to an investigation of the splotch mouse model as an example implementation.
Results:
The protocols provide DTI data in 24 min per direction at 75 microm isotropic using a three-dimensional fast spin-echo sequence, enabling preliminary imaging in 3 h (6 directions plus one unweighted measurement), or detailed imaging in 9 h (42 directions plus six unweighted measurements). Application to the splotch model enabled assessment of spinal cord pathology.
Conclusion:
We present guidelines for designing diffusion microMRI experiments, which may be adapted for different studies and research facilities. As they are suitable for routine use and may be readily implemented, we hope they will be adopted by the phenotyping community.
Insights
New diffusion microscopic MRI (microMRI) protocols accelerate mouse embryo phenotyping. These optimized methods reduce scanning times, enabling routine use for visualizing novel tissue contrast and assessing developmental pathologies.
Area of Science:
- Developmental biology
- Biomedical imaging
- Genetics
Background:
- Advanced imaging techniques are crucial for characterizing mutant mouse embryos.
- Diffusion microscopic MRI (microMRI) offers potential for embryo phenotyping but faces limitations due to long scan times.
- Lack of standardized protocols hinders routine microMRI adoption for embryo analysis.
Purpose of the Study:
- To develop and validate optimized experimental protocols for diffusion microMRI of mouse embryos.
- To address the limitations of extended scanning times in current microMRI procedures.
- To facilitate routine phenotyping of mutant mouse embryos through improved data acquisition.
Main Methods:
- Development of two protocols for diffusion microMRI experimental design in mouse embryos.
- Consideration of embryo preparation and pulse sequence parameters for optimal data.
- Application of protocols to the splotch mouse model for validation.
Main Results:
- Protocols yield Diffusion Tensor Imaging (DTI) data at 75 microm isotropic resolution in 24 min/direction.
- Preliminary imaging achievable in 3 hours, with detailed imaging in 9 hours.
- Successful assessment of spinal cord pathology in the splotch mouse model.
Conclusions:
- Guidelines for designing diffusion microMRI experiments are presented.
- Protocols are adaptable for various research settings and studies.
- The developed methods are suitable for routine use and encourage adoption by the phenotyping community.
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