流式细胞术是一种通过测量激光照射细胞时产生的散射光和荧光来从细胞中获得各种信息的技术。专门设计用于流式细胞术的仪器被称为流式细胞仪。
流式细胞仪依靠特异性抗体的荧光标记来检测细胞内外的标记物,这允许基于细胞类型、活动状态和细胞周期状态等多个参数对细胞群体进行定量评估。
流式细胞仪包含 3 个主要组件:液流系统、光学系统和电子系统。每个组件都扮演着不同的角色。
当用荧光染料标记的细胞悬浮液倒入细管中时,细胞以一定的间隔移动通过细管。
然后用激光光束照射这些细胞。从荧光物质发射出的荧光和由细胞散射的光由光电检测器检测和监测。
1. 靶细胞被荧光标记。
2. 将细胞倒入流路系统中,并以固定的间隔移动通过管。
3. 用激光光束照射细胞。
4. 通过光学系统检测光。
为了获得迄今为止流式细胞术中尚未获得的更多信息,有必要用多种类型的标记物对细胞进行染色。在精确区分各种标记物组合的测量中,利用不同波长的激光器和探测器进行多色分析发挥着积极作用。
传统的荧光检测测量通常限于荧光强度高的波长范围。但是,测量和分析技术的最新进展让我们能够获得整个光谱的荧光信息。这提高了多色分析的精度,并将细胞分析的标准提高到更先进的水平。
在多色分析过程中,由于在测量中使用了多种荧光染料,不同波长的荧光可能在测量项目之间重叠,由于光泄漏到最初不用于测量的通道中而导致干扰。纠正这个问题的技术称为补偿。为了进行补偿,必须预测目标荧光染料之间的荧光重叠并将其从信号中去除。这种补偿处理的精度在很大程度上取决于流式细胞仪中探测器的灵敏度特性。
与多色分析相比,传统流式细胞术不能一次测量尽可能多的项目,但波长光谱之间的重叠较少,这使得测量更容易执行。
由于一次要测量的项目数量很大,这会导致测量光谱之间的重叠,因此多色分析需要复杂而精确的测量。
流式细胞术中的多色分析现在需要具有更高性能的多功能探测器。
为了获得大量的精确数据,必须将检测波长范围扩展到近红外区域,这意味着探测器需要在近红外区域具有更高的灵敏度。
为了满足这些需求,滨松正在努力提高其用于多色分析的探测器性能。
例如,与在光阴极面中使用碱性材料的传统光电倍增管相比,我们不断提高利用光阴极面中晶体材料的光电倍增管模块性能,并实现了更高的灵敏度和扩展的近红外灵敏度。
我们提供一系列光电倍增管,在单个元件中具有多个检测通道,还提供易于使用的模块,其功能非常适合多色分析。
MPPC 模块是带有内置 MPPC 的光子计数模块。此类模块集成了 MPPC、I/V 转换器和电源,因而使用非常方便。
MPPC 具有高增益、高光子探测效率和宽动态范围,因此,即使在高速、弱信号下,也能获得高信噪比特性。
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