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Three-dimensional NMR microscopy of zebrafish specimens
Timothy L Kline1, Caroline R Sussman2, Maria V Irazabal2
1Department of Radiology, Mayo Clinic, Rochester, MN, USA.
NMR in Biomedicine
|November 16, 2018
Summary
Nuclear Magnetic Resonance (NMR) microscopy enables detailed imaging of adult zebrafish, overcoming limitations of optical methods due to pigmentation and growth. This technique allows for high-resolution morphological analysis of zebrafish mutants.
Area of Science:
- Biomedical Imaging
- Zebrafish Model Organisms
- Nuclear Magnetic Resonance
Background:
- Zebrafish embryos are optically transparent, facilitating imaging, but pigmentation and tissue growth in juveniles and adults impede optical analysis.
- Advanced imaging techniques are necessary for studying adult zebrafish morphology and development.
- Nuclear Magnetic Resonance (NMR) is a powerful tool for biological system studies and has been applied to in vivo zebrafish microscopy.
Purpose of the Study:
- To assess the utility of Nuclear Magnetic Resonance microscopy (MRM) for imaging various types of zebrafish specimens.
- To optimize 3D MRM parameters for high-resolution imaging of zebrafish organs.
- To analyze contrast mechanisms and develop segmentation methods for morphological studies of zebrafish mutants.
Main Methods:
- Utilized 3D MRM to image formalin-fixed (FF), formalin-fixed and paraffin-embedded (FFPE), fresh (unfixed), and gadolinium-doped FF zebrafish specimens.
- Acquired 3D NMR images with isotropic resolutions of 50 μm/pixel in 10 minutes and 25 μm/pixel in 4 hours at 7 Tesla.
- Investigated contrast sources, identifying T1-weighting (3D FLASH, 3D FISP) for FF specimens and T2-weighting (3D RARE) for FFPE specimens.
Main Results:
- Demonstrated high-resolution 3D MRM imaging of zebrafish specimens across different fixation and embedding states.
- Achieved isotropic resolutions of 50 μm/pixel within 10 minutes and 25 μm/pixel within 4 hours.
- Determined optimal contrast weighting (T1 for FF, T2 for FFPE) for delineating organ structures.
Conclusions:
- NMR microscopy is a viable method for high-contrast, high-resolution imaging of adult zebrafish, including fixed and fresh specimens.
- Developed and validated MRM protocols for detailed morphological assessment of zebrafish.
- Highlighted the potential for automated analysis of morphological changes in zebrafish mutants using MRM, advancing genetic and developmental studies.
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