Histochemistry on vibratome sections of fish tissue: a comparison of fixation and embedding methods
Histochemistry on vibratome sections of fish tissue: a comparison of fixation and embedding methods
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DOI：
10.3354/ab00624
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发表时间：
2015-01-01
期刊：
影响因子：
1.4
通讯作者：
Harzsch, Steffen
中科院分区：
文献类型：
--
作者：
Grunow, Bianka;Kirchhoff, Tina;Harzsch, Steffen
尽管近年来成像技术有所改进，但对于许多非模型系统，组织固定，然后包埋和切片进行组织化学或免疫组织化学染色仍然是脊椎动物组织学中的一项重要技术。本研究旨在探讨使用不同制备技术保存鱼组织的组织学切片。采用不同固定剂（甲醛、多聚甲醛和甲醛锌）和包埋方法（明胶、琼脂糖和低温熔融琼脂糖）对小斑角鲨（Scyliorhinus canicula）、大西洋鲟（acpenser oxyrinchus）和斑马鱼（Danio rerio）干段的横向振动切片质量进行了比较。我们的数据表明，用于组织化学染色的振动体切片的质量强烈依赖于固定和包埋介质。虽然多聚甲醛固定导致躯干节段比其他固定剂更明显的收缩，但在斑马鱼和角鲨中，该固定剂的切片质量和组织化学染色效果最好。此外，嵌入方法对截面的质量有很大的影响。在角鲨和鲟鱼样本中，首选的包埋介质是琼脂糖和低温融化琼脂糖，因为明胶经常导致组织收缩。总之，对于组织化学检查，振动体切片的处理方案需要单独适应每个研究生物体。
Despite improvements in imaging techniques during recent years, for many non-model systems the fixation of tissues followed by embedding and sectioning for histochemical or immunohistochemical staining remains an important technique in vertebrate histology. The present study sets out to explore the preservation of histological sections of fish tissues using different preparation techniques. The quality of transverse vibratome sections from trunk segments of the lesser-spotted dogfish Scyliorhinus canicula, Atlantic sturgeon Acipenser oxyrinchus and zebrafish Danio rerio were compared using different fixatives (formaldehyde, paraformaldehyde and zinc-formaldehyde) and embedding methods (gelatine, agarose and low-temperature melting agarose). Our data show that the quality of the vibratome sections for histochemical staining is strongly dependent upon fixation and embedding media. Although paraformaldehyde fixation results in a more pronounced shrinkage of the trunk segment than the other fixatives used, the quality of the sections and the histochemical staining was best with this fixative in zebrafish and dogfish. Additionally, the embedding methods have a strong influence on the quality of the sections. In the dogfish and sturgeon samples, the preferred embedding media were agarose and low-temperature melting agarose, since gelatine often caused shrinkage of the tissues. In conclusion, for histochemical examinations, the processing protocols for vibratome sectioning need to be adapted individually to each study organism.