Department of Biosciences, Rice University, Houston, TX 77005, USA.
STAR Protoc. 2021 Jul 30;2(3):100709. doi: 10.1016/j.xpro.2021.100709. eCollection 2021 Sep 17.
Characterizing mRNA and protein expression with temporal and spatial resolution is a valuable component of nearly every developmental study. Here, we describe a protocol that combines in situ hybridization chain reaction (HCR) and immunofluorescence, allowing for the detection of mRNAs and proteins simultaneously, in zebrafish embryos and larvae. This protocol expands the flexibility of multiplexed HCR by coupling it with traditional immunofluorescence detection. For complete details on the use and execution of this protocol, please refer to Choi et al. (2010, 2016, 2018) and Howard et al. (2021).
以时间和空间分辨率来描述 mRNA 和蛋白质的表达是几乎每一项发育研究的一个有价值的组成部分。在这里,我们描述了一个将原位杂交链反应(HCR)和免疫荧光结合起来的方案,允许在斑马鱼胚胎和幼虫中同时检测 mRNAs 和蛋白质。该方案通过将其与传统的免疫荧光检测相结合,扩展了多重 HCR 的灵活性。如需了解本方案的详细使用和执行情况,请参阅 Choi 等人(2010、2016、2018)和 Howard 等人(2021)的研究。