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用于体积电子显微镜的包埋树脂的定量评估。

Quantitative evaluation of embedding resins for volume electron microscopy.

作者信息

Tegethoff Lennart, Briggman Kevin L

机构信息

Max Planck Institute for Neurobiology of Behavior - caesar, Bonn, Germany.

出版信息

Front Neurosci. 2024 Feb 9;18:1286991. doi: 10.3389/fnins.2024.1286991. eCollection 2024.

Abstract

Optimal epoxy resin embedding is crucial for obtaining consistent serial sections from large tissue samples, especially for block faces spanning >1 mm. We report a method to quantify non-uniformity in resin curing using block hardness measurements from block faces. We identify conditions that lead to non-uniform curing as well as a procedure to monitor the hardness of blocks for a wide range of common epoxy resins used for volume electron microscopy. We also assess cutting repeatability and uniformity by quantifying the transverse and sectional cutting forces during ultrathin sectioning using a sample-mounted force sensor. Our findings indicate that screening and optimizing resin formulations is required to achieve the best repeatability in terms of section thickness. Finally, we explore the encapsulation of irregularly shaped tissue samples in a gelatin matrix prior to epoxy resin embedding to yield more uniform sections.

摘要

对于从大型组织样本中获取一致的连续切片而言,最佳的环氧树脂包埋至关重要,特别是对于跨度大于1毫米的块面。我们报告了一种通过测量块面的块硬度来量化树脂固化不均匀性的方法。我们确定了导致固化不均匀的条件以及一种用于监测用于体积电子显微镜的各种常见环氧树脂块硬度的程序。我们还通过使用安装在样品上的力传感器在超薄切片过程中量化横向和切片力来评估切割重复性和均匀性。我们的研究结果表明,需要筛选和优化树脂配方以在切片厚度方面实现最佳重复性。最后,我们探索在环氧树脂包埋之前将不规则形状的组织样本封装在明胶基质中以产生更均匀的切片。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c6e/10884229/e5a012ec5f4a/fnins-18-1286991-g001.jpg

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