Flow-stream waveguide for collection of perpendicular light scatter in flow cytometry

✍ Scribed by Raymond Mariella Jr.; Gerrit van den Engh; Donald Masquelier; Gerald Eveleth

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 482 KB
• Volume: 24
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/(sici)1097-0320(19960501)24:1<27::aid-cyto3>3.0.co;2-f

✦ Synopsis

We report a new physical configuration for the detection of perpendicular light scatter or fluorescence in flow cytometry when using a fluid stream in air. "his configuration increases the signal-tonoise ratio and narrows the coeEicient of variation for uniformly sized latex spheres when compared to using a microscope objective to collect such light. The new technique views the scattered light that is trapped within the optical waveguide that is M ~Urally formed by the flow stream in air. One efficient and simple way to detect the light trapped within this optical waveguide is to place one end of a fiber optic, with a conically polished tip and sufficiently large-core diameter, directly into the flow stream and to place an optical detector at the fiber's other end. For perpendicular light scatter, the flowstream waveguide achieves high collection efficiency (NA = 0.88) as well as high efficiency of optical throughput due to lack of surfaces between the light scatterem and the detector. We obtained 10fold higher signals with this technique than with a long-working-distance microscope objective. The flow-stream waveguide is also much easier to align than traditional microscope-lens-based systems.