Mellen Nicholas M
Kosair Children's Hospital Research Institute, University of Louisville, Louisville, KY 40202, USA.
J Neurosci Methods. 2008 Feb 15;168(1):113-8. doi: 10.1016/j.jneumeth.2007.09.027. Epub 2007 Oct 2.
Slice preparations isolate functional networks, permitting single unit recording under visual control, and the use of fluorescent indicators. Circuits of interest often lie at a tilt in both the rostrocaudal and ventrodorsal axis, thus exposing circuits of interest at the cut surface of a slice would require a device for tilting a preparation along two orthogonal axes relative to the blade. Such a device, designed to be used in conjunction with a vibrating microtome, permitting the isolation of slice preparations at reproducible angles, is described here. Because the two orthogonal axes of tilt can be independently and continuously adjusted, it is possible to use this device to successively refine tilt parameters from preparation to preparation for optimal exposure of circuits of interest, facilitating the development of new slice preparations. Its use in cutting a thick medullary slab preparation, isolated from the neonate rat, which exposes respiratory networks at the cut surface is described.
脑片制备可分离功能网络,允许在视觉控制下进行单神经元记录,并使用荧光指示剂。感兴趣的神经回路通常在前后轴和腹背轴上呈倾斜状态,因此要在脑片的切割面上暴露感兴趣的神经回路,就需要一种能使标本相对于刀片沿两个正交轴倾斜的装置。本文介绍了一种与振动切片机配合使用的装置,该装置能以可重复的角度分离脑片标本。由于两个正交倾斜轴可以独立且连续地调节,因此可以使用该装置在逐个标本的过程中不断优化倾斜参数,以实现对感兴趣神经回路的最佳暴露,从而有助于开发新的脑片标本。文中还描述了该装置在切割从新生大鼠分离出的厚髓板标本中的应用,该标本在切割面上可暴露呼吸网络。