权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development and its application of simple methods for bone sections

简易骨切片方法的开发及其应用

基本信息

批准号：
03557076
负责人：
YOSHIKI Shusaku
金额：
$ 5.12万
依托单位：
Showa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03557076/
关键词：
bone tissue cyanuric chloride decakcified section hitochemistry immunohistochemistry in situ hybridization 骨標識 in situ hybridization

项目摘要

Dr.Yoshiki, a principal investigator of this research project, reported the simple method to identify osteoid tissues in decalcified bone sections. In this method, treatment of bone tissues with cyanuric chloride before decalcification is essential to identify osteoid tissues. In the present study, we conducted several experiments to develop new application of this technique for bone research, and obtained the following results.1) When tartrate acid resistant acid phosphatase (TRACP) activity was detected in various tissues prepared by conventional technique, its activity was found is not only osteoclasts but also mononuclear cells in spleen and lungs. However, its activity was detected in only osteoclasts when tissues were prepared after cyanuric chloride treatment. These results indicate that this technique is useful to identify osteoclast cell lineage in decalcified bone sections.2) We recently developed in situ hybridization technique for bone sections. We examined the effects of cyanuric chloride treatment on in situ hybridization. The expression of mRNAs for osteocalcin and type I collagen is detected in bone sections prepared after cyanuric chloride treatment as well as bone sections prepared by conventional technique. These results suggest that cyanuric chloride treatment dose not affect detection of mRNA by in situ hybridization. This also suggests the possibility of combined technique of histochemistry and in situ hybridization.3) We tried immunohistochemistry using bone sections treated with cyanuric chloride, but we obtained strong background staining. This technique is not useful for immunohistochemistry.

该研究项目的首席研究员yoshiki博士报告了在脱钙骨切片中识别类骨组织的简单方法。在这种方法中，在脱钙化之前用三聚氰胺处理骨组织对于识别类骨组织至关重要。在本研究中，我们进行了多次实验，以开发该技术在骨研究中的新应用，并获得了以下结果。1)测定酒石酸耐酸磷酸酶（TRACP）在常规方法制备的各种组织中的活性，发现其活性不仅存在于破骨细胞中，还存在于脾脏和肺部的单核细胞中。然而，当三聚氰尿酸处理后制备的组织仅在破骨细胞中检测到其活性。这些结果表明，该技术可用于鉴定脱钙骨切片中的破骨细胞谱系。我们最近开发了骨切片原位杂交技术。我们考察了三聚氰胺处理对原位杂交的影响。在三氰氯处理后的骨切片和常规方法制备的骨切片中检测骨钙素和I型胶原mrna的表达。这些结果表明，三聚氰尿酸处理不影响mRNA的原位杂交检测。这也提示了组织化学与原位杂交相结合技术的可能性。3)我们尝试用三聚氰胺处理的骨切片进行免疫组织化学，但我们获得了强背景染色。该技术不适用于免疫组织化学。