Yang Xu, Beniash Elia
Department of Oral Biology, School of Dental Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
Department of Bioengineering, Center for Craniofacial Regeneration, McGowan Institute for Regenerative Medicine, Swanson School of Engineering, University of Pittsburgh, Pittsburgh, PA, USA.
Methods Mol Biol. 2019;1922:191-196. doi: 10.1007/978-1-4939-9012-2_19.
Immunofluorescence (IF) labeling is a powerful technique that can provide a wealth of information on structural organization, supramolecular composition, and functional properties of cells and tissues. At the same time, nonspecific staining and false positives can seriously compromise IF studies and lead to confusing or even misleading results. It is particularly true for the extracellular matrix component of forming enamel. Here, we present an optimized IF protocol for developing enamel. Autofluorescence blocking by Sudan Black B (SBB) and establishing of proper isotype controls lead to a significant artifact reduction and improve reliability of the IF data.
免疫荧光（IF）标记是一种强大的技术，可提供有关细胞和组织的结构组织、超分子组成及功能特性的丰富信息。同时，非特异性染色和假阳性会严重影响IF研究，并导致结果混淆甚至产生误导。对于正在形成的釉质的细胞外基质成分而言尤其如此。在此，我们展示了一种用于发育中釉质的优化IF方案。用苏丹黑B（SBB）阻断自发荧光并建立适当的同型对照可显著减少假象并提高IF数据的可靠性。
