To demonstrate the selective nature of optical sorting, we isolated a subpopulation of macrophages highly infected with the fluorescently labeled pathogen Francisella tularensis subsp. novicida. A total of 10,738 infected cells were sorted at a throughput of 11 cells/s with 93% purity and 39% recovery.
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Isolation of a subpopulation of macrophages highly infected by Alexa—488 labeled F. tularensis novicida. Off-chip FACScan flow cytometry data for: initial mixture of infected and uninfected macrophages (red); waste channel (green); and collection channel (blue). A total of 10,738 cells were sorted at a throughput of 11 cells/s with 93% purity and 39% recovery. |
Infection of mouse macrophages by Alexa-488 labeled F. tularensis novicida. A) 3D reconstruction of stacked confocal images showing internalized as well as surface-adherent bacteria. B) Broad fluorescence histogram taken by FACScan flow cytometer indicating variability in the number of pathogens per macrophage. C) Variability in the number of pathogens (green) per macrophage shown by fluorescence microscopy. |
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Perroud, T. D.; Kaiser, J. N.; Sy, J. C.; Lane, T. W.; Branda, C. S.; Singh, A. K.; Patel, K. D. Microfluidic-Based Cell Sorting of Francisella tularensis Infected Macrophages using Optical Forces. Analytical Chemistry 80, 6365-6372 (2008).