Steinkamp J A, Habbersett R C, Stewart C C
Cytometry. 1987 Jul;8(4):353-65. doi: 10.1002/cyto.990080403.
A modular detector for measuring multicolor fluorescence from cells illuminated by single or multiple lasers has been developed for flow cytometers. Motion picture projector, camera, and CCTV/video lenses were evaluated for use in the detector by comparing their physical characteristics, image quality, and light collection efficiencies. A 25-mm focal length F/0.95 CCTV lens was selected, based on our criteria and test results. The detector was constructed out of square aluminum extrusion channels. A CCTV lens mounted on the outside of the first channel collected light emitted from cells and collimated it towards filters and secondary CCTV lenses located in each channel. The secondary lenses functioned as relay optics for directing and focusing light onto pinhole spatial filters for measurement by photomultipliers. The detector design allowed any number of channels to be connected together and the versatility for making simultaneous or sequential measurements. Measurements on lymphocytes labeled with four monoclonal antibodies conjugated to fluorescent dyes and measurements on viable tumor cells stained for DNA content and with three fluorescent-labeled antibodies were used to demonstrate the detector's capabilities.
一种用于流式细胞仪的模块化探测器已被开发出来，用于测量由单束或多束激光照射的细胞发出的多色荧光。通过比较电影放映机、相机和闭路电视/视频镜头的物理特性、图像质量和光收集效率，对它们在探测器中的使用进行了评估。根据我们的标准和测试结果，选择了一个焦距为25毫米、F/0.95的闭路电视镜头。探测器由方形铝挤压通道制成。安装在第一个通道外部的闭路电视镜头收集细胞发出的光，并将其准直朝向位于每个通道中的滤光片和二级闭路电视镜头。二级镜头用作中继光学器件，将光引导并聚焦到针孔空间滤光片上，以便由光电倍增管进行测量。探测器的设计允许任意数量的通道连接在一起，并具有进行同时或顺序测量的通用性。对用与荧光染料偶联的四种单克隆抗体标记的淋巴细胞进行的测量，以及对用三种荧光标记抗体染色以测定DNA含量的活肿瘤细胞进行的测量，被用来证明探测器的能力。
