A modified tape transfer approach for rapidly preparing high-quality cryosections of undecalcified adult rodent bones.
A modified tape transfer approach for rapidly preparing high-quality cryosections of undecalcified adult rodent bones.
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一种改进的胶带转移方法，用于快速制备未脱钙成年啮齿动物骨骼的高质量冷冻切片
DOI：
10.1016/j.jot.2020.03.001
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发表时间：
2021-01
影响因子：
6.6
通讯作者：
Hong D
中科院分区：
文献类型：
--
作者：
Yang Y;Liu Q;Zhang L;Fu X;Chen J;Hong D
基于组织学的分析是解剖骨骼稳态、疾病和再生的细胞和分子机制的重要工具。这些努力的成功高度依赖于快速获得适合各种分析的矿化骨骼组织的高质量切片。然而，目前用于制备此类切片的技术仍远不能令人满意。本研究的目的是开发一种新的方法来制备高质量的不脱钙骨切片适用于各种组织学分析。对传统的Cryojane胶带转移系统进行了两项重要的修改，包括利用优化的粘合剂来制备粘合剂载玻片以提高转移效率，以及使用廉价的传统台式UV透射仪进行UV固化。使用这种改良的胶带转移方法制备未脱钙啮齿动物骨的冷冻切片，并目视检查其组织形态和结构完整性。对这些切片进行了各种组织学分析，包括钙黄绿素标记、Von kossa染色、免疫荧光和酶活性染色以及5-乙炔基-2 '-脱氧尿苷（EdU）和TUNEL测定。我们开发了一种改进的胶带转移方法，可以在4天内以低成本制备不脱钙啮齿动物成年骨的冷冻切片。通过这种方法制备的骨切片显示出良好的组织形态和结构完整性。此外，这些切片适用于各种组织学分析，包括钙黄绿素标记、Von kossa染色、免疫荧光和酶活性染色以及EdU和TUNEL测定。我们开发的胶带转移方法提供了一种快速，经济实惠，易于学习的方法，用于制备对骨骼研究有价值的高质量不脱钙骨切片。我们的研究提供了一种快速，经济实惠，易于学习的方法，用于制备高质量的未脱钙骨切片，可用于准确诊断各种骨疾病，并评估不同疗法治疗这些疾病的疗效。
Histology-based analyses are important tools to dissect cellular and molecular mechanisms of skeletal homeostasis, diseases, and regeneration. The success of these efforts is highly dependent on rapidly obtaining high-quality sections of mineralized skeletal tissues suitable for various analyses. However, the current techniques for preparing such sections are still far from satisfactory. This study aimed to develop a new approach for preparing high-quality undecalcified bone sections applicable to various histological analyses. Two important modifications were made to the conventional Cryojane Tape-Transfer System, including utilization of an optimized adhesive to prepare adhesive glass slides for improving the transfer efficiency, and a cheap conventional benchtop UV transilluminator for UV curing. Cryosections of undecalcified rodent bones were prepared using this modified tape transfer approach, and their tissue morphology and structural integrity were visually examined. A variety of histological analyses, including calcein labeling, Von kossa staining, immunofluorescence, and enzymatic activity staining as well as 5-Ethynyl-2’-deoxyuridine (EdU) and TUNEL assays, were performed on these sections. We developed a modified version of tape transfer approach that can prepare cryosections of undecalcified rodent adult bones within 4 days at a low cost. Bone sections prepared by this approach exhibited good tissue morphology and structural integrity. Moreover, these sections were applicable to a variety of histological analyses, including calcein labeling, Von kossa staining, immunofluorescence, and enzymatic activity staining as well as EdU and TUNEL assays. The tape transfer approach we developed provides a rapid, affordable, and easy learning method for preparing high-quality undecalcified bone sections valuable for bone research. Our research provides a rapid, affordable, and easy learning method for preparing high-quality undecalcified bone sections that can be potentially used for accurate diagnosis of various bone disorders and evaluation of the efficacy of different therapies in the treatment of these diseases.
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影响因子：
--
作者：
Hootman, JM;Helmick, CG
通讯作者：
Helmick, CG
骨细胞固有的 mTORC1 信号传导抑制小鼠小梁骨的生长
影响因子：
4
作者：
Liu, Qingbai;Liu, Cunchang;Chen, Jianquan
通讯作者：
Chen, Jianquan
影响因子：
6.2
作者：
Chen, Jianquan;Long, Fanxin
通讯作者：
Long, Fanxin
DOI：
10.1007/978-1-62703-989-5_11
发表时间：
2014-01-01
期刊：
SKELETAL DEVELOPMENT AND REPAIR: METHODS AND PROTOCOLS
影响因子：
--
作者：
Kawamoto, Tadafumi;Kawamoto, Komei
通讯作者：
Kawamoto, Komei
DOI：
10.1083/jcb.119.3.493
发表时间：
1992-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Gavrieli Y;Sherman Y;Ben-Sasson SA
通讯作者：
Ben-Sasson SA