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Confirmatory measurement channels for LIF-based bioaerosol instrumentation

Bisson, Scott E.; Crocker, Robert W.; Kulp, Thomas J.; Reichardt, Thomas A.; Reilly, Peter T.A.; Whitten, William B.

As part of the U.S. Department of Homeland Security Detect-to-Protect (DTP) program, a multilab [Sandia National Laboratories (SNL), Lawrence Livermore National Laboratories (LLNL), Pacific Northwest National Laboratory (PNNL), Oak Ridge National Laboratory (ORNL), and Los Alamos National Laboratory (LANL)] effort is addressing the need for useable detect-to-warn bioaerosol sensors for public facility protection. Towards this end, the SNL team is investigating the use of rapid fluorogenic staining to infer the protein content of bioaerosols. This is being implemented in a flow cytometer wherein each particle detected generates coincident signals of correlated forward scatter, side scatter, and fluorescence. Several thousand such coincident signal sets are typically collected to generate a distribution describing the probability of observing a particle with certain scattering and fluorescence values. These data are collected for sample particles in both a stained and unstained state. A linear unmixing analysis is performed to differentiate components in the mixture. In this paper, we discuss the implementation of the staining process and the cytometric measurement, the results of their application to the analysis of known and blind samples, and a potential instrumental implementations that would use staining.