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We tackled this matter by utilizing rotating-polarization coherent anti-Stokes Raman scattering (RP-CARS) microscopy to review the effects various fixatives in the myelin sub-micrometric molecular purchase and micrometric morphology. RP-CARS is a novel strategy based on VEHICLES microscopy enabling probing spatial orientation of molecular bonds while maintaining the intrinsic chemical selectivity of VEHICLES microscopy. By characterizing the results for the fixation processes, the present work signifies a helpful guid