Flow Cytometric Analysis of Murine Microbiota in T Cell Transfer Colitis 1

In recent years, high-resolution analysis of bacterial forward scatter and DNA content by flow cytometry has enabled the near-realtime analysis of dynamic changes in the composition of complex bacterial ecosystems (Koch C et al., Nature Protocols, 2013). We sought to apply this method to detect changes in the composition of the fecal microbiota in mice in the course of experimentally induced inflammation of the colon.
Conclusion:
Measuring bacterial forward scatter and DNA content, we were able to discriminate up to 80 sub-populations in fecal microbiomes of mice before and after the onset of colitis. Our method identified dramatic differences in the composition of the fecal microbiota in colitis compared to healthy mice. Fluorescent beads FluoSpheres 2 µm (505/515, Lot-Nr. 24498W, Molecular Probes Eugene, Oregon, USA), FluoSpheres 1 µm (350/440, Lot-Nr. 69A1-1, Molecular Probes Eugene, Oregon, USA), and Fluoresbrite BB Carboxylate microspheres 0.5 µm (360/407, Lot-Nr. 552744, Polyscience, USA) were used to align the instrumental settings.