Journal: bioRxiv

Article Title: Motility Functions as an Essential Defense in the Bacterial Persister Lifecycle

doi: 10.64898/2025.12.17.694804

Figure Lengend Snippet: A) Schematic of generating rifampin-induced persisters and evaluating persister survival rates. B) Survival rate (mean value ± standard deviation) of E. coli MG1655 cells against ampicillin with no pretreatment ( A ) and rifampin-pretreatment ( R-A ). Data represent results from six independent experiments. C) Motile percentage (mean value ± standard deviation) of mid-exponential phase cells ( M ), rifampin-treated cells ( R ), rifampin-induced persisters ( R-A ). Data represent results from five experimental videos. The total numbers of cells in the five videos are 598 ( M ), 663 ( R ), and 291 ( R-A ), respectively. D - F) Representative trajectories of mid-exponential phase cells ( D ), rifampin-treated cells ( E ), and persisters ( F ) swimming in their original living environment (LB or LB with corresponding antibiotics), imaged in the FCS2 chamber. The colored lines show the trajectories of different cells. G , H) Comparison of track mean speed ( G ) and track mean directional change rate ( H ) distributions for mid-exponential phase cells ( M ), rifampin-treated cells ( R ), and persisters ( R-A ). Data were obtained from at least three experimental videos. Sample sizes for mid-exponential phase cell, rifampin-treated cell, and persister are 6358, 6580, and 4610 tracks, respectively.

Article Snippet: To capture the natural swimming behavior of cells in their original living environment, we directly placed 8 μL of bacterial culture onto the Bioptechs FCS2 chamber (0.2 mm plastic pad between slide and coverslip) and used a Nikon ECLIPSE Ti inverted microscope to record bright-field videos at approximately 22 frames per second (FPS).

Techniques: Standard Deviation, Comparison

Journal: bioRxiv

Article Title: Motility Functions as an Essential Defense in the Bacterial Persister Lifecycle

doi: 10.64898/2025.12.17.694804

Figure Lengend Snippet: A) Representative trajectories of mid-exponential phase cells swimming in LB after CCCP exposure, observed in the FCS2 chamber. The colored lines show the trajectories of different cells. B) Survival rate (mean value ± standard deviation) of E. coli cells against ampicillin with exposure to CCCP before rifampin ( C-R-A ), exposure to CCCP and rifampin together ( CR-A ), exposure to CCCP and ampicillin together after rifampin ( R-CA ), and 15% w/v Ficoll in culture medium upon rifampin exposure ( 15%FR-A ). Data represent results from six independent experiments. C) Membrane potential (mean value ± standard deviation) of mid-exponential phase cells treated with CCCP ( CCCP ), experiencing no treatment ( M ), treated with 5% ( 5%F ), 10% ( 10%F ), and 15% ( 15%F ) w/v Ficoll, as indicated by the red/green median fluorescence intensity ratio measured by flow cytometry. Data represent results from at least three independent experiments. D) Representative trajectories of mid-exponential phase cells swimming in LB with 15% w/v Ficoll, imaged in the FCS2 chamber. The colored lines show the trajectories of different cells. E) Two-dimensional distribution correlating track mean speed and track mean directional change rate for mid-exponential phase cells in LB with 15% w/v Ficoll, with corresponding marginal distributions. The sample size is 5374 tracks, obtained from seven videos. F) Relationship between rifampin-induced survival rate against ampicillin (mean) and swimming speed (mean) for cells under different conditions: M , 15%F , and CCCP .

Article Snippet: To capture the natural swimming behavior of cells in their original living environment, we directly placed 8 μL of bacterial culture onto the Bioptechs FCS2 chamber (0.2 mm plastic pad between slide and coverslip) and used a Nikon ECLIPSE Ti inverted microscope to record bright-field videos at approximately 22 frames per second (FPS).

Techniques: Standard Deviation, Membrane, Fluorescence, Flow Cytometry