Citation: Want, A. ... Et Al, 2011. Multi-parameter Ow Cytometry and Cell Sorting Reveal Extensive Physiological Heterogeneity in Bacillus Cereus Batch Multi-parameter Flow Cytometry and Cell Sorting Reveal Extreme

Here we tried to develop a standard flow cytometry protocol for measuring the physiological state of the Gram +‟ve bacterium Bacillus cereus, and attempted to understand more of the resulting staining phenomena using cell sorting. Specifically the use of DiOC6(3) and the proprietary RedoxSensor Green TM (an indicator of bacterial reductase activity) both counter stained with propidium iodide. The data presented here has identified at least 4 distinguishable physiological states based on the two staining protocols alone and dependent on the position in the growth cycle single cells gave rise to varying numbers of colonies when sorted individually onto nutrient agar plates. Further that these growing colonies derived from single cell sorts had widely different lag phases. This highlights further the unexpectedly complex population dynamics of bacterial cultures and lays to rest the microbiological dogma that all bacterial cells in a culture behave in exactly the same way since they are clearly members of a physiologically heterogenous and dynamic population.