Almubarak Asra, Berry Fred B
Department of Medical Genetics, University of Alberta, Edmonton, AB, Canada.
Department of Surgery, University of Alberta, Edmonton, AB, Canada.
Methods Mol Biol. 2022;2579:227-233. doi: 10.1007/978-1-0716-2736-5_17.
Cell proliferation is one of the key events that regulates organism development. In the limb, chondrocytes differentiate into a multi-layered cellular template called the growth plate. Chondrocyte proliferation is essential to provide the necessary cells that allow growth of a bone. Deregulated cell proliferation will lead to truncated bone elements. Immunofluorescence is a biological technique that uses specific antibodies to detect the subcellular localization of a proliferative marker within cellular or tissue context. In this chapter, we illustrate how to perform immunofluorescence to detect the localization of Ki-67 (a marker of actively growing/proliferating chondrocytes) in order to assess the growth fraction of the columnar chondrocytes in the growth plate in paraffin-embedded mouse tissue limb.
细胞增殖是调节机体发育的关键事件之一。在四肢中，软骨细胞分化形成一种称为生长板的多层细胞模板。软骨细胞增殖对于提供使骨骼生长所需的细胞至关重要。细胞增殖失控会导致骨元素发育不全。免疫荧光是一种生物技术，它利用特异性抗体在细胞或组织环境中检测增殖标志物的亚细胞定位。在本章中，我们将说明如何进行免疫荧光检测，以检测Ki-67（活跃生长/增殖软骨细胞的标志物）的定位，从而评估石蜡包埋的小鼠四肢组织生长板中柱状软骨细胞的生长分数。
