Bailey Charles G, Rasko John E J
Gene and Stem Cell Therapy Program, Centenary Institute of Cancer Medicine and Cell Biology, Australia.
Methods Mol Biol. 2007;411:99-110. doi: 10.1007/978-1-59745-549-7_7.
The unique spectral properties and versatility of autofluorescent proteins have facilitated their widespread use in flow cytometric applications. The ability to analyze heterologous fluorescent protein expression conveniently and noninvasively by individually interrogating cells has facilitated increasingly more sophisticated experimental designs to address important biological questions. Improved multilaser flow cytometers have allowed the fluorescent protein field to flourish by permitting high-speed, multiparametric analysis of biological samples. Fluorescent proteins are well suited for either transient or stable expression analysis. Therefore, achieving efficient gene transfer and expression in cells by transfection or viral transduction is paramount to the optimal use of fluorescent proteins in flow cytometry. The archetypal autofluorescent protein, enhanced green fluorescent protein (eGFP), can be used successfully in combination with other fluorescent protein variants. Two such variants, Cerianthus sp. orange fluorescent protein (cOFP) and a fast maturing variant of Discosoma sp. red protein (DsREDExpress), are well suited for flow cytometric applications in combination with eGFP and do not require special filters for optimal excitation and detection.
自发荧光蛋白独特的光谱特性和多功能性促进了它们在流式细胞术应用中的广泛使用。通过逐个检测细胞来方便且无创地分析异源荧光蛋白表达的能力，推动了越来越复杂的实验设计，以解决重要的生物学问题。改进的多激光流式细胞仪通过对生物样品进行高速多参数分析，让荧光蛋白领域蓬勃发展。荧光蛋白非常适合瞬时或稳定表达分析。因此，通过转染或病毒转导在细胞中实现高效基因转移和表达，对于在流式细胞术中最佳使用荧光蛋白至关重要。典型的自发荧光蛋白——增强型绿色荧光蛋白（eGFP），可以与其他荧光蛋白变体成功联用。两种这样的变体，Cerianthus sp.橙色荧光蛋白（cOFP）和盘状珊瑚红色蛋白（DsREDExpress）的快速成熟变体，非常适合与eGFP联用进行流式细胞术应用，并且不需要特殊滤光片来实现最佳激发和检测。
