Summary:
The HET-FLOW project aims to develop high-resolution cell biology tools using flow cytometry to simultaneously detect transcripts and proteins, and possibly other functional characteristics such as membrane potential, in the same cell.
We'll be using the Lactococcus cremoris model, a fermentative bacterium that nevertheless possesses respiratory chain elements, one of which, cytochrome bd, exhibits production heterogeneity from one cell to another under certain environmental conditions. To determine whether this protein heterogeneity is linked to heterogeneity at transcriptional level, we will transpose the flow-FISH method already established for eukaryotic cells to bacterial cells. We will also use fluorescent markers of membrane potential to determine whether heterogeneity in protein production translates into functional heterogeneity, testing the respiratory capacity of cells. The ultimate goal will be to correlate protein production, gene expression and respiratory chain functionality at the single-cell level.
Once we have perfected our model, the tools we develop will be made available to the microbiology community, enabling them to study phenotypic variability and bacterial adaptation strategies.