Preparation of Thin Frozen Sections from Nonfixed and Undecalcified Hard Tissues Using Kawamot's Film Method (2012)
Preparation of Thin Frozen Sections from Nonfixed and Undecalcified Hard Tissues Using Kawamot's Film Method (2012)
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DOI：
10.1007/978-1-62703-989-5_11
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发表时间：
2014-01-01
期刊：
影响因子：
--
通讯作者：
Kawamoto, Komei
中科院分区：
文献类型：
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作者：
Kawamoto, Tadafumi;Kawamoto, Komei
描述了一种通过使用粘合膜制备硬组织切片的方法。该方法从成年小鼠和大鼠的骨骼中产生非常薄（至1微米厚）的冷冻切片。将骨组织用水溶性介质冷冻包埋，然后在切割表面上安装粘合膜后用一次性碳化钨刀片切割。将切片染色在粘合膜上，并保存在粘合膜和载玻片之间。包埋、切割、染色、封片等步骤仅用20分钟即可完成，软硬组织保存完好，骨髓保存完好。细胞如成骨细胞、成纤维细胞和破骨细胞被清楚地识别，并且骨的类骨质层被清楚地观察到。切片适用于多种类型的染色，如组织学、组织化学、酶组织化学、免疫组织化学和原位杂交。免疫组化可用不固定和不脱钙切片进行。除了这些应用之外，切片还用于观察PGF荧光。切片也可用于研究组织中水溶性物质的分布。此外，切片对于使用LMD技术的基因分析和成像质谱非常有用。
A method for preparing hard tissue sections by using an adhesive film is described. The method produces very thin (to one-micrometer thick) frozen sections from adult mouse and rat bone. The bone tissue is freeze-embedded with water-soluble medium and then cut with a disposable tungsten carbide blade after mounting the adhesive film onto the cut surface. The sections are stained on the adhesive film and preserved between the adhesive film and glass slide. All the steps including the embedding, cutting, staining, and mounting are completed within only 20 min. The soft and hard tissues are preserved satisfactorily and the bone marrow is also preserved perfectly. Cells such as osteoblasts, fibroblasts, and osteoclasts are clearly identified, and the osteoid layer of bone is clearly observed. The sections are applicable to many types of staining such as histology, histochemistry, enzyme histochemistry, immunohistochemistry, and in situ hybridization. The immunohistochemistry can be carried out with nonfixed and undecalcified sections. In addition to these applications, the sections are used for observing the PGF fluorescence. The sections are also usable for studying the distribution of water-soluble materials in the tissues. Furthermore, the sections are very useful for gene analysis using LMD technique and for imaging mass spectrometry.