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The reliability of the challenge test depends, among other parameters, on the spatial distribution of microorganisms in the matrix. The present study aims to quickly identify factors that are susceptible to impair a uniform distribution of inoculated bacteria in cosmetic matrices in this context. We used mosaic confocal laser scanning microscopy (M-CLSM) to obtain rapid assessment of the impact of the composition and viscosity of cosmetic matrices on S. aureus spatial distribution. Severaldoi:10.3390/cosmetics7020032 fatcat:o6tuh52w7bczdnrfcxkqulynfy