Characterization of the sensitivity of side scatter in a flow‐stream waveguide flow cytometer

• Published in: Cytometry (New York, N.Y.) N.Y.), 1999-10, Vol.37 (2), p.160-163
• Main Authors: Mariella, Raymond P., Huang, Zhengping, Langlois, Richard G.
• Format: Article
• Language: English
• Subjects: flow cytometry; Flow Cytometry - instrumentation; Flow Cytometry - methods; flow‐stream waveguide; sensitivity; Sensitivity and Specificity; side scatter
• ISSN: 0196-4763; 1097-0320
• DOI: 10.1002/(SICI)1097-0320(19991001)37:2<160::AID-CYTO10>3.0.CO;2-3

Background: We previously reported a new optical configuration, in which both the side scatter and the fluorescence are collected using the index‐guided, total internal reflection of a flow stream in air (the flow‐stream waveguide). Methods: Using a mixture of 0.202‐μm and 0.093‐μm diameter polystyrene beads, we have characterized the side scatter (SSC) sensitivity of a custom‐built flow cytometer (miniFlo) which incorporates a flow‐stream waveguide. Results: The SSC‐triggered SSC signal of 0.093‐μm polystyrene beads in water was almost baseline resolved from the background. We also measured the SSC‐triggered SSC signal of the same beads in water on our FACScan, which is a commercial unit with the conventional optical arrangement that uses a custom imaging objective to collect light from a sheath flow cuvette in perpendicular direction—the signal from 0.093‐μm beads was not resolved from the background. Conclusions: The SSC sensitivity of miniFlo is one of the best reported in the literature. Cytometry 37:160–163, 1999. Published 1999 Wiley‐Liss, Inc.