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Micro-Optical Sectioning Tomography to Obtain a High-Resolution Atlas of the Mouse Brain
Existing imaging tools have limitations for brainwide mapping of neural circuits at a mesoscale level. In collaboration with DiATOME, researchers developed a Micro-Optical Sectioning Tomography (MOST) system utilizing a DiATOME Diamond Knife that can provide micron tomography of a centimeter-sized whole mouse brain.
Slicing was performed by moving the specimen to generate ribbons, and each ribbon was simultaneously imaged. The illuminating beam passed through a beam splitter, mirror and objective to irradiate the ribbon. The imaging beam collected by the objective and passed through the mirror, beam splitter and tube lens was then recorded by a line-scan CCD.
A 3D structural dataset of a Golgi-stained whole mouse brain at the neurite level was obtained. The morphology and spatial locations of neurons and traces of neurites were clearly distinguished. Researchers found that neighboring Purkinje cells were sticking to each other.
Creating a High Resolution Atlas of the Mouse Brain... (A) A sagittal image reconstructed from a stack of 100 virtual sagittal sections (total thickness of 0.1 mm). These sections were transformed from the original coronal sections. The sagittal image was located in the right hemisphere about 0.4 mm lateral to the middle. Almost all major regions of the brain can be seen in this image, e.g., the Olfactory Bulb (OB), Cerebral Cortex (Cx), Hippocampus (Hc), Fornix(f), Anterior Commissure (ac), Thalamus (T), Cerebellum (Cb), Midbrain (Mb), Pons (P), Medulla (Md), Corpus Callosum (cc), Superior Colliculus (SC), Inferior Colliculus (IC), Hypothalamus (Ht), Preoptic Area (Po), Optic Chiasm (ox), 4th ventricle (4V) and nine lobules of the cerebellum (Arabic numerals, 2 to 10). The three regions inside the different colored rectangle in (A) are the positions of (B), (C) and (D), which illustrate the cerebral cortex, hippocampus and cerebellum, respectively. In the reconstruction of sagittal image, no dislocation was observed along the D-V axis, i.e., the coronal sections are inherently aligned along the A-P axis.
Acknowledgement Micro-Optical Sectioning Tomography to Obtain a High-Resolution
Atlas of the Mouse Brain Anan Li, Hui Gong, Bin Zhang, Qingdi Wang, Cheng Yan, Jingpeng Wu, Qian Liu, Shaoqun Zeng, Qingming Luo
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics–Huazhong University of Science and Technology, Wuhan 430074, P. R. China.
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