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Mid-infrared photo-induced force microscopy (PiF-IR) on model system Bacillus subtilis - vancomycin

2025/05/14 by Maryam Ali, Ali, Maryam, Robin Schneider +25
Biochemistry, Genetics and Molecular Biology · Chemistry · Engineering · #Biological Physics (physics.bio-ph) #FOS: Physical sciences #Photoacoustic and Ultrasonic Imaging #Soft Condensed Matter (cond-mat.soft) #Spectroscopy Techniques in Biomedical and Chemical Research #thermodynamics and calorimetric analyses

paper · pdf · doi:10.48550/arxiv.2505.10249

openalex publication_date 2025/05/14 · openalex created_date 2025/10/11 · openalex updated_date 2026/07/28

Abstract

The power of photothermal spectroscopic imaging to visualize antimicrobial interaction on the surface of individual bacteria cells has been demonstrated on the model system Bacillus subtilis and vancomycin using mid-infrared photo-induced force microscopy (PiF-IR, also mid-IR PiFM). High-resolution PiF contrasts obtained by merging subsequent PiF-IR scans at two different illumination frequencies revealed chemical details of cell wall destruction after 30 and 60 min incubation with vancomycin with a spatial resolution of ≈ 5 nm. This approach compensates local intensity variations induced by near-field coupling of the illuminating electric field with nanostructured surfaces, which appear in single-frequency contrasts in photothermal imaging methods, as shown by [Anindo et al., J. Phys. Chem C, 2025, 129, 4517]. Known spectral shifts associated with hydrogen bond formation between vancomycin and the N-acyl-D-Ala4-D-Ala5 termini in the peptidoglycan cell wall have been observed in chemometrics of PiF-IR spectra from treated and untreated Bacillus subtilis harvested after 30 min from the same experiment. Spectral signatures of the vancomyin interaction have been located in the piecrust of a progressing septum with ≈ 10 nm resolution using PiF contrasts of three selected bands of a PiF-IR hyperspectral scan of an individual Bacillus subtilis cell harvested after 30 min incubation. Our results are complemented by a discussion of imaging artifacts and the influence of parameter settings supporting further development towards standardization in the application of PiF-IR for visualizing the chemical interaction of antibiotics on the surface of microbes with few nanometer resolution.

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