Asquith Becca, Debacq Christophe, Florins Arnaud, Gillet Nicolas, Sanchez-Alcaraz Teresa, Mosley Angelina, Willems Luc
Department of Immunology, Imperial College London, Norfolk Place, London W2 1PG, UK.
Proc Biol Sci. 2006 May 7;273(1590):1165-71. doi: 10.1098/rspb.2005.3432.
The cytoplasmic dye carboxyfluorescein diacetate succinimidyl ester (CFSE) is used to quantify cell kinetics. It is particularly important in studies of lymphocyte homeostasis where its labelling of cells irrespective of their stage in the cell cycle makes it preferable to deuterated glucose and BrdU, which only label dividing cells and thus produce unrepresentative results. In the past, experiments have been limited by the need to obtain a clear separation of CFSE peaks forcing scientists to adopt a strategy of in vitro labelling of cells followed by their injection into the host. Here we develop a framework for analysis of in vivo CFSE labelling data. This enables us to estimate the rate of proliferation and death of lymphocytes in situ, and thus represents a considerable advance over current procedures. We illustrate this approach using in vivo CFSE labelling of B lymphocytes in sheep.
细胞质染料羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）用于量化细胞动力学。在淋巴细胞稳态研究中，它尤为重要，因为它对细胞进行标记时不考虑其在细胞周期中的阶段，这使其比仅标记分裂细胞从而产生不具代表性结果的氘代葡萄糖和溴脱氧尿苷更具优势。过去，实验受到需要清晰分离CFSE峰的限制，迫使科学家采用先在体外标记细胞然后将其注入宿主的策略。在此，我们开发了一个用于分析体内CFSE标记数据的框架。这使我们能够原位估计淋巴细胞的增殖和死亡速率，因此相对于当前方法有了相当大的进步。我们通过对绵羊B淋巴细胞进行体内CFSE标记来说明这种方法。
