Hashimoto Hisashi, Kusakabe Moriaki, Ishikawa Hiroshi
Department of Anatomy, The Jikei University School of Medicine, Minatoku, Tokyo 105-8461, Japan.
Microsc Res Tech. 2008 Jan;71(1):51-9. doi: 10.1002/jemt.20522.
A novel method for acquiring serial images suitable for three-dimensional reconstruction of vascular networks in the whole brain of mouse was developed. The brain infused with a White India ink-gelatin solution was fixed and embedded in paraffin containing Sudan Black B through xylene also containing Sudan Black B. Each sliced surface of the paraffin block was coated with liquid paraffin and its image was serially acquired. Coating with liquid paraffin extremely improved the quality of the image. The series of serial images was free of distortion and a three-dimensional image was reconstructed without the problem of the alignment and registration of adjacent images. The volume-rendered image indicated three-dimensional distribution of blood vessels in a whole brain. No ghost or shadow was observed on a volume-rendered image of the White India ink-gelatin infused brain. The z-axial resolution examined on the orthogonal sections reconstituted from serial images obtained at an interval of 5 mum showed no cross talk, indicating that the z-axial resolution was no larger than 5 mum. A proper understanding of the vascular system in a whole brain is indispensable to reveal the development of the vascular system in the brain of normal and genetically manipulated mouse and vascular alterations in pathological situation, such as stroke and neurodegenerative disease. Although simple and inexpensive, this method will provide fundamental information on the vascular system in a whole brain.
开发了一种新方法，用于获取适合对小鼠全脑血管网络进行三维重建的系列图像。将注入白色印度墨水 - 明胶溶液的大脑固定，通过同样含有苏丹黑B的二甲苯嵌入含有苏丹黑B的石蜡中。石蜡块的每个切片表面都涂有液体石蜡，并对其图像进行连续采集。涂覆液体石蜡极大地提高了图像质量。该系列连续图像无失真，无需对相邻图像进行对齐和配准即可重建三维图像。体绘制图像显示了全脑中血管的三维分布。在注入白色印度墨水 - 明胶的大脑的体绘制图像上未观察到重影或阴影。从以5μm间隔获得的系列图像重建的正交切片上检查的z轴分辨率显示无串扰，表明z轴分辨率不大于5μm。正确了解全脑中的血管系统对于揭示正常和基因操作小鼠脑中血管系统的发育以及病理情况下（如中风和神经退行性疾病）的血管改变是必不可少的。尽管该方法简单且成本低廉，但它将提供关于全脑中血管系统的基础信息。
