The optical density of each culture at 620nm was measured with a Thermo Scientific Multiskan FC microplate spectrophotometer, following the end of each growth period27 (link). To measure the fractions of each population, at the end of each growth period a portion of cells from each well (typically 5 μL, unless the low density conditions made it necessary to transfer larger volumes) was transferred to a new plate containing 200μL sterile Cellgro PBS buffer (Mediatech, VA). These samples were then analyzed at a BD LSRIIHTS flow cytometer operating in high throughput (HT) mode, where we determined the ratio between the producer and freeloader strains, distinguished by their fluorescence emissions 36 (link). Given the total density and the strain ratio, the densities of the producer and freeloader populations were readily calculated.
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Measuring Microbial Population Dynamics
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Other organizations : Massachusetts Institute of Technology, Harvard University
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Variable analysis
independent variables
- Growth period duration
- Optical density at 620 nm
- Fraction of producer and freeloader strains in the population
- Volume of cell samples transferred to the new plate (typically 5 μL, unless the low density conditions made it necessary to transfer larger volumes)
- Sterile Cellgro PBS buffer used to dilute the cell samples
- BD LSRIIHTS flow cytometer operating in high throughput (HT) mode
- Positive control: Not specified
- Negative control: Not specified
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