Data for paper - Autofluorescence-Based Detection of Antibiotic Resistance by Spectral Flow Cytometry

<p style="text-align:justify; margin-bottom:8px">Flow cytometry in microbiology faces challenges such as limited bacterial-specific stains, high fluorophore costs, poor stain permeability, and difficulty distinguishing small bacterial cells from debris. Staining also requires multiple wash steps that lower sensitivity and add complexity. Autofluorescence detection, linked to metabolic cofactors like FAD and NADH, offers a stain-free, cost-effective alternative. We present a method to distinguish antibiotic-resistant from non-resistant bacteria using autofluorescence spectral signatures captured with a spectral flow cytometer (seven lasers, 54 detectors, and an integrated biosafety cabinet). Gentamicin-stressed <em data-end="787" data-start="778">E. coli</em> and <em data-end="804" data-start="792">Salmonella</em> showed increased autofluorescence at UV, violet, and blue wavelengths compared with controls. Similarly, oxacillin-stressed MSSA exhibited elevated autofluorescence within 4–6 hours, while MRSA showed no significant change. These results demonstrate that antibiotic resistance can be rapidly and quantitatively detected through resistance-specific autofluorescence, offering a promising stain-free diagnostic tool.</p>