Utilizing decellularized bio-membranes to optimize histopathological embedding of small tissues

IF 1.6 Q2 MULTIDISCIPLINARY SCIENCES MethodsX Pub Date : 2024-08-20 DOI:10.1016/j.mex.2024.102919
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Abstract

In recent years, minimally invasive biopsy techniques have been widely used to generate small tissue samples that require processing in clinical pathology. However, small paraffin-embedded tissues are prone to loss due to their small size. To prevent the loss of small tissues, researchers have employed nonbiological embedding materials for preembedding, but this approach can lead to cumbersome experimental procedures and increase the chances of tissue loss. This study aimed to develop a convenient decellularized embedding material derived from biological membrane tissues to effectively protect small tissues from loss during paraffin embedding. This study decellularized three types of fresh animal-derived membrane tissues and selected the small intestine as the most suitable decellularized raw material through attempts at softening, comparing physical properties, and using tissue as the starting material. Subsequently, small tissues from various tissue sources were embedded, followed by H&E staining, Masson staining, immunofluorescence staining, and immunohistochemical staining. The decellularized material derived from biomembrane tissues (DMBT) developed in this study can reduce the loss of small tissues without the need for preembedding, thereby shortening the embedding process. This provides a new pathological embedding tool for future laboratory and clinical research and work.

  • The fat layer of the pig's small intestine is scraped off, and chemical reagents are used to defat and decellularize it.

  • Chemical reagents are used to soften and make the pig's small intestine transparent, and the decellularized pig's small intestine is dried.

  • DMBT is used for embedding and staining the biological tissue.

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利用脱细胞生物膜优化小组织的组织病理学包埋
近年来,微创活检技术已被广泛应用于产生需要在临床病理学中处理的小组织样本。然而,石蜡包埋的小组织由于体积小,容易丢失。为了防止小型组织的丢失,研究人员采用了非生物包埋材料进行预包埋,但这种方法会导致实验过程繁琐,增加组织丢失的几率。本研究旨在开发一种源自生物膜组织的便捷脱细胞包埋材料,以有效保护小组织在石蜡包埋过程中免受损失。本研究对三种新鲜动物来源的膜组织进行脱细胞处理,并通过软化、比较物理性质和以组织为起始材料等尝试,选择小肠作为最合适的脱细胞原料。随后,对不同组织来源的小组织进行包埋,然后进行 H&E 染色、Masson 染色、免疫荧光染色和免疫组化染色。本研究开发的生物膜组织脱细胞材料(DMBT)无需预包埋即可减少小组织的损失,从而缩短了包埋过程。刮掉猪小肠的脂肪层,用化学试剂对其进行脱脂和脱细胞处理,用化学试剂软化猪小肠并使其透明,然后将脱细胞的猪小肠干燥,用 DMBT 对生物组织进行包埋和染色。
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来源期刊
MethodsX
MethodsX Health Professions-Medical Laboratory Technology
CiteScore
3.60
自引率
5.30%
发文量
314
审稿时长
7 weeks
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