Yosuke Otake

Quantitative Histological Validation of Diffusion Tensor MRI with Two-Photon Microscopy of Cleared Mouse Brain

1Department of Radiology, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan 2Research Institute for Diseases of Old Age, Juntendo University Graduate School of Medicine 3Research & Development Group, Hitachi Ltd. 4Department of Radiology, University of Tokyo 5Department of Radiological Sciences, Graduate School of Human Health Sciences 6Department of Neuropathology, Tokyo Medical and Dental University (Received November 22, 2015; Accepted February 10, 2016; published online March 30, 2016)

See-through Brains and Diffusion Tensor MRI Clarified Fiber Connections: A Preliminary Microstructural Study in a Mouse with Callosal Agenesis

Clearing methods that render the brain optically transparent allow high-resolution three-dimensional (3D) imaging of neural networks. We used diffusion tensor imaging (DTI) and two-photon imaging of cleared brains to analyze white matter in BTBR mice. We confirmed corpus callosum agenesis and identified an abnormal commissure close to the third ventricle. DTI and cleared-brain two-photon imaging revealed that these commissural fibers constituted a frontal clustering of the ventral hippocampal commissure and provided a detailed assessment of white matter structure in mice.

Magnetic Resonance Microscopy of Chemically Fixed Human Embryos at High Spatial Resolution

We acquired magnetic resonance (MR) microscopic images of chemically fixed human embryos of Carnegie stages 16 to 22 with a large image matrix (256 × 256 × 512) using an MR microscope that we developed with a 9.4-tesla vertical wide-bore superconducting magnet and a dual-channel receiver system to extend the dynamic range of the MR signal. The images showed clear anatomical structures at spatial resolutions of (40 µm)(3) to (60 µm)(3). We concluded that the experimental technique we developed will aid construction of the next anatomical database of the collection of chemically fixed human embryos.

Diffusion-weighted Line-scan Echo-planar Spectroscopic Imaging Technique to Reduce Motion Artifacts in Metabolite Diffusion Imaging

Metabolite diffusion is expected to provide more specific microstructural and functional information than water diffusion. However, highly accurate measurement techniques have still not been developed, especially for reducing motion artifacts caused by cardiac pulsation and respiration. We developed a diffusion-weighted line-scan echo-planar spectroscopic imaging (DW-LSEPSI) technique to reduce such motion artifacts in measuring diffusion-weighted images (DWI) of metabolites. Our technique uses line-scan and echo-planar techniques to reduce phase errors induced by such motion during diffusion time. The phase errors are corrected using residual water signals in water suppression for each acquisition and at each spatial pixel specified by combining the line-scan and echo-planar techniques. We apply this technique to a moving phantom and a rat brain in vivo to demonstrate the reduction of motion artifacts in DWI and apparent diffusion coefficient (ADC) maps of metabolites. DW-LSEPSI will be useful for investigating a cellular diffusion environment using metabolites as probes.

The Relationship between Neurite Density Measured with Confocal Microscopy in a Cleared Mouse Brain and Metrics Obtained from Diffusion Tensor and Diffusion Kurtosis Imaging

Purpose: Diffusional kurtosis imaging (DKI) enables sensitive measurement of tissue microstructure by quantifying the non-Gaussian diffusion of water. Although DKI is widely applied in many situations, histological correlation with DKI analysis is lacking. The purpose of this study was to determine the relationship between DKI metrics and neurite density measured using confocal microscopy of a cleared mouse brain. Methods: One thy-1 yellow fluorescent protein 16 mouse was deeply anesthetized and perfusion fixation was performed. The brain was carefully dissected out and whole-brain MRI was performed using a 7T animal MRI system. DKI and diffusion tensor imaging (DTI) data were obtained. After the MRI scan, brain sections were prepared and then cleared using aminoalcohols (CUBIC). Confocal microscopy was performed using a two-photon confocal microscope with a laser. Forty-eight ROIs were set on the caudate putamen, seven ROIs on the anterior commissure, and seven ROIs on the ventral hippocampal commissure on the confocal microscopic image and a corresponding MR image. In each ROI, histological neurite density and the metrics of DKI and DTI were calculated. The correlations between diffusion metrics and neurite density were analyzed using Pearson correlation coefficient analysis. Results: Mean kurtosis (MK) (P = 5.2 × 10−9, r = 0.73) and radial kurtosis (P = 2.3 × 10−9, r = 0.74) strongly correlated with neurite density in the caudate putamen. The correlation between fractional anisotropy (FA) and neurite density was moderate (P = 0.0030, r = 0.42). In the anterior commissure and the ventral hippocampal commissure, neurite density and FA are very strongly correlated (P = 1.3 × 10−5, r = 0.90). MK in these areas were very high value and showed no significant correlation (P = 0.48). Conclusion: DKI accurately reflected neurite density in the area with crossing fibers, potentially allowing evaluation of complex microstructures.