Pioli KimAnh T, Pioli Peter D
Department of Biochemistry, Microbiology and Immunology, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N5E5, Canada.
STAR Protoc. 2023 May 12;4(2):102308. doi: 10.1016/j.xpro.2023.102308.
Antibody-secreting cells (ASCs) are critical regulators of the humoral immune response. However, differences between tissue resident populations versus those that have recently migrated to their final anatomic destination are poorly understood. Here, we present a protocol for using retro-orbital (r.o.) CD45 antibody labeling to identify tissue resident versus recently immigrated ASCs in mice. We describe steps for r.o. injection of antibodies, animal euthanasia, and tissue harvesting. We then detail tissue processing, cell counting, and cell staining for flow cytometry analysis. For complete details on the use and execution of this protocol, please refer to Pioli et al. (2023)..
抗体分泌细胞（ASC）是体液免疫反应的关键调节因子。然而，组织驻留群体与最近迁移到其最终解剖位置的群体之间的差异尚不清楚。在这里，我们提出了一种使用眶后（r.o.）CD45抗体标记来识别小鼠组织驻留与最近迁入的ASC的方案。我们描述了眶后注射抗体、动物安乐死和组织采集的步骤。然后，我们详细介绍了组织处理、细胞计数和用于流式细胞术分析的细胞染色。有关本方案使用和执行的完整详细信息，请参阅皮奥利等人（2023年）的文章。
