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Myelin content is an important marker for neuropathology and MRI generated myelin water fraction (MWF) has been shown to correlate well with myelin content. However, because MWF is based on the amount of signal from myelin water, that is, the water trapped between the myelin lipid bilayers, the reading may depend heavily on myelin morphology. This is of special concern when there is a mix of intact myelin and myelin debris, as in the case of injury. To investigate what MWF measures in the presence of debris, we compared MWF to transmission electron microscopy (TEM) derived myelin fraction that measures the amount of compact appearing myelin. A rat spinal cord injury model was used with time points at normal (normal myelin), 3 weeks post-injury (myelin debris), and 8 weeks post-injury (myelin debris, partially cleared). The myelin period between normal and 3 or 8 weeks post-injury cords did not differ significantly, suggesting that as long as the bilayer structure is intact, myelin debris has the same water content as intact myelin. The MWF also correlated strongly with the TEM-derived myelin fraction, suggesting that MWF measures the amount of compact appearing myelin in both intact myelin and myelin debris. From the TEM images, it appears that as myelin degenerates, it tends to form large watery spaces within the myelin sheaths that are not classified as myelin water. The results presented in this study improve our understanding and allows for better interpretation of MWF in the presence of myelin debris.Myelin water fraction (MWF) was validated with high-resolution electron microscopy.Compact appearing intact myelin and myelin debris were quantified.Myelin content and myelin periodicity were measured.No significant change in periodicity was found in compact appearing debris.MWF correlated highly to compact myelin content in intact myelin and myelin debris.