A new instrument using reflectance transformation imaging (RTI), named MorphoLight, has been developed for surface characterization. This instrument is designed to be adjustable to surfaces, ergonomic, and uses a combination of high-resolution imaging functions, i.e., focus stacking (FS) and high dynamic range (HDR), to improve the image quality. A topographical analysis method is proposed with the instrument. This method is an improvement of the surface gradient characterization by light reflectance (SGCLR) method. This aims to analyze slope/curvature maps, traditionally studied in RTI, but also to find the most relevant lighting position and 3D surface parameter which highlight morphological signatures on surfaces and/or discriminate surfaces. RTI measurements and analyses are performed on two zones, sky and sea, of a naval painting which have the same color palette but different painting strokes. From the statistical analysis using bootstrapping and analysis of variance (ANOVA), it is highlighted that the high-resolution images (stacked and tonemapped from HDR images) improve the image quality and make it possible to better see a difference between both painting zones. This difference is highlighted by the fractal dimension for a lighting position (θ, φ) = (30°, 225°); the fractal dimension of the sea part is higher because of the presence of larger brushstrokes and painting heaps.