| 窦丹丹 董玲凤 文华 李自立 胡亮.HistoGel预包埋法在微小组织石蜡切片中的应用[J].,2015,15(28):5405-5407 |
| HistoGel预包埋法在微小组织石蜡切片中的应用 |
| Application of HistoGel Pre-embedding for Preparing Paraffin Sections ofMicrotissue |
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| DOI： |
| 中文关键词: HistoGel 微小组织 石蜡切片 |
| 英文关键词: HistoGel Microtissue Paraffin sections |
| 基金项目:国家自然科学基金项目（81372627）；湖南省自然科学基金项目（13JJ338）；
中南大学研究生自主探索创新基金项目（72150050335） |
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| 中文摘要: |
| 目的：探讨微小组织HistoGel预包埋的石蜡切片制作方法和优势。方法：选择拟胚体(embryoid body, EB)为微小组织，将人
类胚胎干细胞(human Embryonic Stem Cell, hESC)切割成小块，用拟胚体培养基在低贴附培养皿中悬浮培养形成拟胚体。收集拟
胚体，4 %的多聚甲醛固定。将HistoGel 加热到60 ℃熔解，离心去除拟胚体中的固定液，把液态的HistoGel 加到拟胚体上，调整拟
胚体的相对位置使其相对集中，待胶冷却凝固，将含有拟胚体的胶块取出，进行常规的石蜡切片操作，包括脱水、透明、浸蜡、包
埋、切片和苏木素- 伊红(hematoxylin-eosin, HE)染色。显微镜下观察拟胚体的形态，并用拟胚体的形态完好度来评判这种方法。结
果：HistoGel 仅在60 ℃便可熔解，室温可以冷却凝固，含有拟胚体的胶块在整个操作过程中很方便。展片过程中，石蜡和HistoGel
能够保持平整。HE 染色的结果可以看出，拟胚体内部结构完好，细胞核和细胞质清晰可辨。结论：HistoGel可以作为一种微量细
胞组织的预包埋胶制作石蜡切片，而且相比琼脂预包埋，HistoGel 因其操作更加方便和特有的物理特性显示出更多的优点。 |
| 英文摘要: |
| Objective:To investigate the method and the advantages of preparing paraffin sections of microtissues with HistoGel
embedding.Methods:Embryoid body (EB) was chosed to be an example of microtissues. Human Embryonic Stem Cell (hESC) clones
were cut into small pieces, suspended in EB medium, cultured in low-attachment tissue culture dishes to form EBs. The embryoid bodies
were collected and fixed in 4 % paraformaldehyde. HistoGel was heated into liquid state at 60 ℃ upon use. After centrifugation and
removing the supernatant fixatives, the fixed EBs were mixed with prewarmed liquid HistoGel and adjusted to relatively condensed state.
The mixture was cooled down to get solid state at room temperature. Following the solidification, the gel was routinely processed to
check the histology. Briefly, the solidified mixture was dehydrated, paraffin embedded, sectioned, and stained with hematoxylin-eosin.
The sections were observed under microscope. The morphology of the EBs was checked to evaluate the method.Results:HistoGel could
be easily heated to liquid state at only 60 ℃ and cooled down to solid state at room temperature. The solidified mixture could be easily
processed under regular histological protocols. The paraffin and HistoGel mixture section spread well in water bath. The morphology of
the EBs under microscope was well preserved. Additionally, infrastructure with clear nuclei detail and intact cytoplasmwas also detected
clearly.Conclusion:HistoGel double embedding method could be a good choice for microtissue histological analysis because of its
convenience. Moreover, HistoGel showed more benefits than agarose embedding because of its convenience and unique characteristic. |
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