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

✍ Scribed by Raymond P. Mariella Jr; Zhengping Huang; Richard G. Langlois

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 134 KB
• Volume: 37
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/(sici)1097-0320(19991001)37:2<160::aid-cyto10>3.0.co;2-3

✦ Synopsis

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 directionthe 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.