Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, USA.
J Neurosci Methods. 2011 Jul 15;199(1):78-81. doi: 10.1016/j.jneumeth.2011.03.027. Epub 2011 Apr 8.
This paper presents a simple method to measure tissue slice thicknesses using an ohmmeter. The circuit described here is composed of a metal probe, an ohmmeter, a counter electrode, culture medium or physiological buffer, and tissue slice. The probe and the electrode are on opposite interfaces of an organotypic hippocampal slice culture. The circuit closes when the metal probe makes contact with the surface of the tissue slice. The probe position is recorded and compared to its position when it makes contact with the insert membrane on which the tissue grows, thus yielding a thickness measurement. The method does not reduce the viability of slice cultures. Thicknesses of the slice cultures were measured under a number of culturing protocols. An initial drop in thickness occurred between 0 and 4 days in culture. Thicknesses are rather constant thereafter. The type of culture medium and the initial thickness of the tissue explant influence the thickness. Slice thicknesses were compared to a known technique by using optical measurements of slice cross-sections to obtain thicknesses. In contrast to this known technique, the proposed method does not sacrifice the slice culture for measurement purposes. The proposed measurement technique described is straightforward and rapid, about 1 min per culture.
本文提出了一种使用欧姆计测量组织切片厚度的简单方法。这里描述的电路由金属探头、欧姆计、对电极、培养基或生理缓冲液和组织切片组成。探头和电极位于器官型海马切片培养物的相对界面上。当金属探头与组织切片表面接触时，电路闭合。记录探头的位置，并将其与组织生长的插入膜接触时的位置进行比较，从而得出厚度测量值。该方法不会降低切片培养物的活力。在多种培养方案下测量了切片培养物的厚度。在培养的 0 到 4 天之间，厚度会初始下降。此后，厚度相当稳定。培养介质的类型和组织外植体的初始厚度会影响厚度。通过对切片横截面进行光学测量来获得厚度，将切片厚度与已知技术进行了比较。与已知技术相比，该方法不会为了测量目的而牺牲切片培养物。所描述的测量技术简单快速，每个培养物约需 1 分钟。
