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采用振动切片机获取的猪和鼠脑组织切片的蛋白质组分析。

Profile analysis of hepatic porcine and murine brain tissue slices obtained with a vibratome.

机构信息

Research Center "E. Piaggio," University of Pisa , Pisa , Italy.

Research Center "E. Piaggio," University of Pisa , Pisa , Italy ; Institute of Clinical Physiology, National Research Council , Pisa , Italy.

出版信息

PeerJ. 2015 Apr 30;3:e932. doi: 10.7717/peerj.932. eCollection 2015.

Abstract

This study is aimed at characterizing soft tissue slices using a vibratome. In particular, the effect of two sectioning parameters (i.e., step size and sectioning speed) on resultant slice thickness was investigated for fresh porcine liver as well as for paraformaldehyde-fixed (PFA-fixed) and fresh murine brain. A simple framework for embedding, sectioning and imaging the slices was established to derive their thickness, which was evaluated through a purposely developed graphical user interface. Sectioning speed and step size had little effect on the thickness of fresh liver slices. Conversely, the thickness of PFA-fixed murine brain slices was found to be dependent on the step size, but not on the sectioning speed. In view of these results, fresh brain tissue was sliced varying the step size only, which was found to have a significant effect on resultant slice thickness. Although precision-cut slices (i.e., with regular thickness) were obtained for all the tissues, slice accuracy (defined as the match between the nominal step size chosen and the actual slice thickness obtained) was found to increase with tissue stiffness from fresh liver to PFA-fixed brain. This quantitative investigation can be very helpful for establishing the most suitable slicing setup for a given tissue.

摘要

本研究旨在使用振动切片机对软组织切片进行特征描述。特别地,研究了两种切片参数(即步长和切片速度)对新鲜猪肝以及多聚甲醛固定(PFA 固定)和新鲜鼠脑的切片厚度的影响。建立了一个简单的切片包埋、切片和成像框架,以获取其厚度,并通过专门开发的图形用户界面对其进行评估。切片速度和步长对新鲜肝切片的厚度影响不大。相反,PFA 固定的鼠脑切片的厚度取决于步长,但与切片速度无关。鉴于这些结果,仅改变步长对新鲜脑组织进行切片,结果发现这对所得切片厚度有显著影响。尽管为所有组织获得了精密切割的切片(即厚度规则),但发现切片精度(定义为所选名义步长与实际获得的切片厚度之间的匹配)随着组织硬度从新鲜肝到 PFA 固定脑而增加。这种定量研究对于为给定组织建立最合适的切片设置非常有帮助。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55aa/4419543/f9d20c850ea3/peerj-03-932-g001.jpg

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