Foo Lynette C
Department of Neurobiology, School of Medicine, Stanford University, Stanford, California 94305, USA.
Cold Spring Harb Protoc. 2013 Jun 1;2013(6):551-60. doi: 10.1101/pdb.prot074229.
The purification of astrocytes by fluorescence-activated cell sorting (FACS) requires that an astrocyte-specific promoter drive the expression of the green fluorescent protein (GFP). Our laboratory uses FACS to acutely isolate astrocytes from young and old tissue as well as to isolate GFP-negative neurons at the end of the FACS sorting to conduct comparative unbiased, large-scale gene expression studies. Because of the relatively harsh nature of FACS sorting, few astrocytes or neurons survive long enough after the sort to be cultured.
通过荧光激活细胞分选(FACS)纯化星形胶质细胞需要一个星形胶质细胞特异性启动子来驱动绿色荧光蛋白(GFP)的表达。我们实验室使用FACS从年轻和年老组织中急性分离星形胶质细胞,并在FACS分选结束时分离GFP阴性神经元,以进行比较性、无偏倚的大规模基因表达研究。由于FACS分选的性质相对苛刻,分选后很少有星形胶质细胞或神经元存活足够长的时间用于培养。