Suppr超能文献

在冷冻切片过程中使用市售胶带以保持软骨和骨组织的完整性。

The use of commercially available adhesive tapes to preserve cartilage and bone tissue integrity during cryosectioning.

作者信息

Serowoky Maxwell A, Patel Divya D, Hsieh Jason W, Mariani Francesca V

机构信息

Department of Stem Cell Biology & Regenerative Medicine, Broad Center for Regenerative Medicine & Stem Cell Research, Keck School of Medicine, University of Southern California, CA, USA.

出版信息

Biotechniques. 2018 Oct;65(4):191-196. doi: 10.2144/btn-2018-0021.

Abstract

The use of fluorescent tags to monitor protein expression and to lineage-trace cells has become a standard complement to standard histological techniques in the fields of embryology, pathology and regenerative medicine. Unfortunately, traditional paraffin embedding protocols can substantially diminish or abolish the native emission signal of the fluorophore of interest. To preserve the fluorescent signal, an alternative is to use cryosectioning; however, this can often result in undesirable artefacts such as tearing or shattering - particularly for mineralized tissues such as bone and cartilage. Here we present a method of using a commercially available tape to stabilize murine femur tissue, thus allowing for cryosectioning of cartilage and bone tissues carrying fluorescent tags without the need for demineralization.

摘要

在胚胎学、病理学和再生医学领域,使用荧光标签监测蛋白质表达和追踪细胞谱系已成为标准组织学技术的一种标准补充方法。不幸的是,传统的石蜡包埋方案会大幅减弱或消除感兴趣的荧光团的天然发射信号。为了保留荧光信号,一种替代方法是使用冷冻切片;然而,这常常会导致诸如撕裂或破碎等不良伪像——特别是对于骨和软骨等矿化组织。在这里,我们展示了一种使用市售胶带稳定小鼠股骨组织的方法,从而能够对带有荧光标签的软骨和骨组织进行冷冻切片,而无需脱矿。

相似文献

3
Demineralized Murine Skeletal Histology.
Methods Mol Biol. 2021;2230:283-302. doi: 10.1007/978-1-0716-1028-2_16.
4
5
Cryosectioning.
Methods Mol Biol. 2013;1018:301-11. doi: 10.1007/978-1-62703-444-9_28.
7
Immunofluorescence microscopy for localization of Arabidopsis chloroplast proteins.
Methods Mol Biol. 2011;774:33-58. doi: 10.1007/978-1-61779-234-2_3.
8
Fixation and paraffin-embedding of mouse tissues for GFP visualization.
Cold Spring Harb Protoc. 2009 Oct;2009(10):pdb.prot5298. doi: 10.1101/pdb.prot5298.
9
Studying gene expression in bone by in situ hybridization.
Methods Mol Biol. 2012;816:305-20. doi: 10.1007/978-1-61779-415-5_20.
10
How to prepare lung specimens for sectioning in cryostat.
Biotech Histochem. 2011 Jun;86(3):207-8.

引用本文的文献

2
Histological assessment of microtia cartilage, a potential source of autograft tissue in ear reconstruction.
J Anat. 2024 Aug;245(2):339-345. doi: 10.1111/joa.14044. Epub 2024 Apr 4.
5
A murine model of large-scale bone regeneration reveals a selective requirement for Sonic Hedgehog.
NPJ Regen Med. 2022 May 17;7(1):30. doi: 10.1038/s41536-022-00225-8.
8
Methodology, selection, and integration of fracture healing assessments in mice.
J Orthop Res. 2021 Nov;39(11):2295-2309. doi: 10.1002/jor.25172. Epub 2021 Sep 10.
9
A novel cryo-embedding method for in-depth analysis of craniofacial mini pig bone specimens.
Sci Rep. 2020 Nov 11;10(1):19510. doi: 10.1038/s41598-020-76336-3.
10
Immunostaining of Skeletal Tissues.
Methods Mol Biol. 2021;2221:261-273. doi: 10.1007/978-1-0716-0989-7_15.

本文引用的文献

1
Screening Gene Knockout Mice for Variation in Bone Mass: Analysis by μCT and Histomorphometry.
Curr Osteoporos Rep. 2018 Apr;16(2):77-94. doi: 10.1007/s11914-018-0421-4.
2
High-Throughput, Multi-Image Cryohistology of Mineralized Tissues.
J Vis Exp. 2016 Sep 14(115):54468. doi: 10.3791/54468.
3
Analysis of Mouse Growth Plate Development.
Curr Protoc Mouse Biol. 2016 Mar 1;6(1):67-130. doi: 10.1002/9780470942390.mo150094.
4
A method for fixing and paraffin embedding tissue to retain the natural fluorescence of reporter proteins.
Biotechniques. 2015 Sep 1;59(3):153-5. doi: 10.2144/000114328. eCollection 2015 Sep.
5
Theory and Practice of Lineage Tracing.
Stem Cells. 2015 Nov;33(11):3197-204. doi: 10.1002/stem.2123. Epub 2015 Aug 18.
6
Natural large-scale regeneration of rib cartilage in a mouse model.
J Bone Miner Res. 2015 Feb;30(2):297-308. doi: 10.1002/jbmr.2326.
7
Manual hematoxylin and eosin staining of mouse tissue sections.
Cold Spring Harb Protoc. 2014 Jun 2;2014(6):655-8. doi: 10.1101/pdb.prot073411.
8
Demineralized murine skeletal histology.
Methods Mol Biol. 2014;1130:165-183. doi: 10.1007/978-1-62703-989-5_12.
9
Genetic mouse models for bone studies--strengths and limitations.
Bone. 2011 Dec;49(6):1242-54. doi: 10.1016/j.bone.2011.08.021. Epub 2011 Aug 31.
10
Generation and characterization of Col10a1-mcherry reporter mice.
Genesis. 2011 May;49(5):410-8. doi: 10.1002/dvg.20733.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验