Department of Science and Environment, Roskilde University, Universitetsvej 1, Postbox 260, DK-4000 Roskilde, Denmark.
Novo Nordisk Foundation Center for Stem Cell Biology (Danstem), University of Copenhagen, Blegdamsvej 3B, DK-2200 Copenhagen N, Denmark.
STAR Protoc. 2021 Jun 30;2(3):100636. doi: 10.1016/j.xpro.2021.100636. eCollection 2021 Sep 17.
We here report a flow-cytometry-based protocol to measure single-cell protein expression in small samples. The protocol is optimized for simultaneous detection of fluorescent proteins and intracellular and surface antigens in the embryonic pancreas from the mouse. Owing to low cell numbers, current protocols for flow cytometric analysis of embryonic tissues rely on tissue pooling. Our protocol enables analysis of one pancreas per sample, thereby facilitating detection of biological variation and minimizing the number of experimental animals needed. For complete details on the use and execution of this protocol, please refer to Nyeng et al (2019).
我们在此报告了一种基于流式细胞术的方案,用于测量小样本中单细胞的蛋白表达。该方案针对从小鼠胚胎胰腺中同时检测荧光蛋白以及细胞内和细胞表面抗原进行了优化。由于细胞数量较少,目前用于胚胎组织流式细胞分析的方案依赖于组织混合。我们的方案可以实现每个样本分析一个胰腺,从而有助于检测生物学变异性并最小化所需实验动物的数量。如需详细了解本方案的使用和执行,请参考 Nyeng 等人(2019 年)。