精确切割肝脏切片石蜡包埋的快速方法

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-08-05 DOI:10.1016/j.tice.2024.102511
Juan Manuel Herrera , Paula Viviani , María Victoria Miró , Adrián Luis Lifschitz , Guillermo León Virkel
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引用次数: 0

摘要

精切肝片(PCLS)是一种组织外植体,被广泛用作代谢和毒性研究的体内外模型。在进行体外试验时,必须进行组织形态学评估,作为整个试验过程中活力分析的一部分。优质 PCLS 组织切片很难获得,因为它们难以操作,在处理过程中可能会收缩或折叠。此外,文献中也没有详细介绍所用的包埋工艺。在本文中,我们提出了一种调整后的快速石蜡包埋杂交母牛 PCLS 的方法。每个 PCLS 都用一块纱布覆盖,然后放入一个组织学盒中。将这些盒放入一系列浴槽中:80%乙醇浴 10 分钟;96%乙醇浴 3 次,每次 10 分钟;丁醇浴 3 次,每次 10 分钟;丁醇-石蜡(1:1)浴 1 次,每次 20 分钟;石蜡浴 3 次,每次 20 分钟。使用折叠纸盒制作石蜡块。可以获得完整的切片,细胞形态保存完好,没有伪影,组织外观与之前通过常规方案处理的 PCLS 相似,这表明所采用的方法是适当的。
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Rapid method for paraffin embedding of precision-cut liver slices

Precision-cut liver slices (PCLS) are tissue explants extensively used as an ex vivo model for metabolism and toxicity studies. When in vitro assays are conducted, it is imperative to perform a histomorphological evaluation as part of the viability analyses throughout the assay time. It is considered that good quality PCLS histological sections are difficult to obtain because they are hard to manipulate, and may shrink or fold during processing. Moreover, bibliography is not detailed on the embedding processes used. In this article, we propose an adjusted and rapid method for paraffin embedding of PCLS from crossbreed steers. Each PCLS was covered with a piece of gauze and placed into a histological cassette. These cassettes were submitted to a series of baths: 80 % ethanol for 10 min; 3 baths of 96 % ethanol for 10 min; 3 baths of butanol for 10 min; 1 bath of butanol-paraffin (1:1) for 20 min in a 60 °C laboratory oven; and 3 baths of paraffin for 20 min in a 60 °C laboratory oven. Folded paper boxes were used to produce paraffin blocks. It was possible to obtain complete sections with preserved cell morphology and no artifacts, and tissue appearance was similar to previous PCLS processed through the routine protocol, demonstrating the adequacy of the method implemented.

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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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