Articular cartilage posesses unique material properties due to a complex depth-dependent composition of sub-components. Raman spectroscopy has proven valuable in quantifying this composition through cartilage crosssections. However, cross-sectioning requires tissue destruction and is not practical in situ. In this work, Raman spectroscopybased multivariate curve resolution (MCR) was employed in porcine cartilage samples (n = 12) to measure collagen, glycosaminoglycan, and water distributions through the surface for the first time; these were compared against cross-section standards. Through the surface Raman measurements proved reliable in predicting composition distribution up to a depth of approximately 0.5 mm. A fructose-based optical clearing agent (OCA) was also used in an attempt to further improve depth of resolution of this measurement method. However, it did not; mainly due to a high-spectral overlap with the Raman spectra of main cartilage sub-components. This measurement technique potentially could be used in situ, to better understand the etiology of joint diseases such as osteoarthritis (OA). K E Y W O R D S articular cartilage, multivariate analysis, optical clearing, Raman spectroscopy 1 | INTRODUCTION Articular cartilage serves as the bearing surface for boneto-bone load transmission, and enables nearly frictionfree joint movement. [1] Fulfilling this role requires unique material properties that are derived from its complex composition and organization. Past studies using Raman spectroscopy have reported the relative distribution of this composition in vitro with cross-sectioning, [2,3] and related it to concentrations determined from assay tests. However, quantifying the composition with depth through the surface will keep morphology intact, and enable in situ measurements. This could offer better insight into how joint diseases such as osteoarthritis (OA) develop, especially in the early stages, and how composition affects functionality. Cartilage consists of a dense extracellular matrix (ECM) predominantly made up of collagens, proteoglycans, and water as well as other proteins and molecules in lower concentrations. [1,4] Recently, tools such as Raman spectroscopy have proven capable of not only quantifying these sub-component concentrations, but also observing subtle variations in their structural organization. [5,6]