Journal: bioRxiv
Article Title: Uncovering the multifaceted mechanism of action of a historical antimicrobial
doi: 10.64898/2026.02.02.703249
Figure Lengend Snippet: A. S. aureus Newman, A. baumannii ATCC 19606, and P. aeruginosa PA14 were stained with DiSC3(5), treated with either 1×MIC of Bald’s eyesalve (purple) or water (black), and their fluorescence monitored for 1 h. There was increased DiSC3(5) fluorescence, indicating membrane depolarisation in samples treated with Bald’s eyesalve. The red arrow indicate point of treatment. Each line represents the mean of three technical replicates. B. S. aureus Newman, A. baumannii ATCC 19606, and P. aeruginosa PA14 were treated with 1×MIC of Bald’s eyesalve (purple) or water (grey) for 1 h, stained with propidium iodide, and fluorescence was measured after 1 h. Each symbol represents an individual bacterial culture; box plots show the fluorescence intensity (in RFU) of propidium iodide. C. S. aureus Newman and A. baumannii ATCC 19606 were treated with 2×MIC of Bald’s eyesalve or water for 2 h, stained with FM4-64, and then imaged with a widefield fluorescence microscope at oil immersion lens (x100) magnification. Increased FM4-64 fluorescence in S. aureus and the presence of dense spots of fluorescence in A. baumannii suggest membrane perturbation.
Article Snippet: For S. aureus , all reads were aligned to S. aureus Newman whole genome sequence (accession number - NC009641.1 , assembly - GCF_000010465.1), while A. baumannii reads were aligned to A. baumannii ATCC 19606 whole genome sequence (accession number - NZ_CP046654.1, assembly - GCF_009759685.1).
Techniques: Staining, Fluorescence, Membrane, Microscopy