Exvivo ESR Spin Rrobe Investigation of Stratum Corneum Lipids

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Electron spin resonance (ESR) is useful for studying stratum corneum (SC)lipid ordering. Order parameter obtained from ESR spectra can be a good index for evaluating the fluidity of SC lipids. However, the method for evaluation of ex vivo SC lipids has not been established. We investigated the details of the ESR methods. A glass plate using a cyanoacrylate resin was used to strip the SC lipids. A single-chain aliphatic spin probe, 5-Doxylstearic acid (5-DSA), was used. The SC samples were incubated in the 0.001% 5-DSA in a 0.1% ethanolic aqueous solution for 60 min at 37℃. After the incubation, an ESR glass holder with the incubated SC sample was mounted in the ESR cavity. The newly designed ESR glass holder improved sample preparations as well as ESR measurements. ESR spectra of the SC samples were fully analyzed using a slow-tumbling simulation. The order parameter obtained by the simulation showed depth dependence of SC lipids as a function of depth in the SC. Thus, we established sample preparations of SC lipids and precise ESR measurements and spectral analyses. In addition, we found depth dependence of the ex vivo SC lipids ordering for the first time.