RESUMEN
To date, detailed studies of the thickness of coatings using surface plasmon resonance have been limited to samples that are very uniform in thickness, and this technique has not been applied quantitatively to samples that are inherently rough or undergo instabilities with time. Our manuscript describes a significant improvement to surface plasmon resonance imaging (SPRi) that allows this sensitive technique to be used for quantitative tracking of the thickness and roughness of surface coatings that are rough on the scale of tens of nanometers. We tested this approach by studying samples with an idealized, one-dimensional roughness: patterned channels in a thin polymer film. We find that a novel analysis of the SPRi data collected with the plane of incidence parallel to the patterned channels allows the determination of the thickness profile of the channels in the polymer film, which is in agreement with that measured using atomic force microscopy. We have further validated our approach by performing SPRi measurements perpendicular to the patterned channels, for which the measured SPR curve agrees well with the single SPR curve calculated using the average thickness determined from the thickness profile as determined using AFM. We applied this analysis technique to track the average thickness and RMS roughness of cellulose microfibrils upon exposure to cellulolytic enzymes, providing quantitative determinations of the times of action of the enzymes that are of direct interest to the cellulosic ethanol industry.