Bjarkam C R, Pedersen M, Sørensen J C
Department of Neurobiology, Institute of Anatomy, University of Aarhus, DK-8000 Aarhus C, Denmark.
J Neurosci Methods. 2001 Jul 30;108(2):153-9. doi: 10.1016/s0165-0270(01)00383-1.
We present a newly developed brain slicing machine and technique for tissue embedding, which enable orientation of fresh or fixed brain tissue from small laboratory animals, in any given position, and subsequent tissue sectioning into slabs with an optional thickness between 0.5 and 20 mm. The oriented tissue slabs may be analysed directly, or processed further on a cryostat or vibratome, into thin stainable histological sections, and aligned to MR-images or brain atlases, depending on the reference used for the initial orientation. Additionally, we describe a new embedding medium (HistOmer) which is an alginate cold polymer ready for instant use after mixing with water. HistOmer allows accurate positioning of the tissue during embedding, and at the same time supports and protects the embedded tissue during sectioning. HistOmer is, therefore, described comprehensively and compared with other commonly used embedding media. This novel slicing technique may also, as illustrated, be used to perform isotropic random orientation of the embedded tissue, before sectioning into tissue slabs of the same thickness. The technique thereby fulfills the requirements for optimal tissue sampling and preparation needed for an unbiased stereologic analysis.
我们展示了一种新开发的用于组织包埋的脑切片机和技术，该技术能够将小型实验动物的新鲜或固定脑组织在任何给定位置进行定向，随后将组织切成厚度在0.5至20毫米之间的任意厚度的薄片。定向后的组织薄片可以直接进行分析，或者在低温恒温器或振动切片机上进一步处理成可染色的薄组织学切片，并根据初始定向所使用的参考，与磁共振图像或脑图谱对齐。此外，我们描述了一种新的包埋介质（HistOmer），它是一种藻酸盐冷聚合物，与水混合后即可立即使用。HistOmer在包埋过程中能使组织准确定位，同时在切片过程中支撑和保护包埋的组织。因此，对HistOmer进行了全面描述，并与其他常用的包埋介质进行了比较。如图所示，这种新颖的切片技术还可用于在将包埋组织切成相同厚度的组织薄片之前，对其进行各向同性随机定向。该技术从而满足了无偏立体分析所需的最佳组织采样和制备要求。
