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We have developed a method combining microfluidics, time-lapsedsingle-molecule microscopy and automated image analysis allowing for theobservation of an excess of 3000 complete cell cycles of exponentially growingEscherichia coli cells per experiment. The method makes it possible to analyzethe rate of gene expression at the level of single proteins over the bacterialcell cycle. We also demonstrate that it is possible to count the number ofnon-specifically DNA binding LacI-Venus molecules using short excitation lightpulses. The transcription factors are localized on the nucleoids in the celland appear to be uniformly distributed on chromosomal DNA. An increase of theexpression of LacI is observed at the beginning of the cell cycle, possiblybecause some gene copies are de-repressed as a result of partitioninginequalities at cell division. Finally, observe a size-growth rate uncertaintyrelation where cells living in rich media vary more in the length at birth thanin generation time and the opposite is true for cells living in poorer media.

Hi-throughput gene expression analysis at the level of single proteins using a microfluidic turbidostat and automated cell tracking