Haugland R P
Molecular Probes, Inc. Eugene, Oregon 97402.
Methods Cell Biol. 1994;42 Pt B:641-63. doi: 10.1016/s0091-679x(08)61100-0.
Flow cytometry uses a relatively small set of dyes for immunochemistry and nucleic acid detection, most of which have been known and used reliably in flow cytometry for several years. These can usually be combined to make simultaneous two-color measurements of multiple cell-surface antigens and nucleic acid content. Because of the overlap of dye spectra and difficulties in finding dyes with substantial Stokes shifts that can be excited by the argon laser, simultaneous three-color or more detection can be more difficult. A basic knowledge of the factors that go into producing the fluorescent signal, including the spectra of dyes and their overlap, is necessary in planning multicolor experiments. By contrast, there have been a number of new fluorescent probes developed for detecting ions, membrane potential, metabolism, organelles, and other properties of living cells as well as for determining cell viability, proliferation, and cell tracking. So far most of these physiological probes are being used only for fundamental research rather than for cell classification. However, as research activity expands in this area, its diagnostic potential is likely to be increasingly appreciated.
流式细胞术使用相对较少的一组染料进行免疫化学和核酸检测，其中大多数染料在流式细胞术中已被熟知并可靠使用多年。这些染料通常可以组合起来，同时对多种细胞表面抗原和核酸含量进行双色测量。由于染料光谱的重叠以及难以找到能被氩激光激发且具有较大斯托克斯位移的染料，同时进行三色或更多色检测可能会更加困难。在规划多色实验时，有必要对产生荧光信号的因素有基本的了解，包括染料的光谱及其重叠情况。相比之下，已经开发出了许多新的荧光探针，用于检测离子、膜电位、代谢、细胞器以及活细胞的其他特性，还用于确定细胞活力、增殖和细胞追踪。到目前为止，这些生理探针大多仅用于基础研究，而非细胞分类。然而，随着该领域研究活动的扩展，其诊断潜力可能会越来越受到重视。
