Serowoky Maxwell A, Patel Divya D, Hsieh Jason W, Mariani Francesca V
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.
Biotechniques. 2018 Oct;65(4):191-196. doi: 10.2144/btn-2018-0021.
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.
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。