Centrifugation of a homogenate of white matter, in a solution of buffered sucrose containing salt, produces a floating layer of myelinated axons. When these are suspended in hypotonic buffer, the mnyelin swells and strips away from the axon. Axons are then separated from the myelin by centrifugation. The resulting preparation consists of a variable population of processes with lengths up to 200 micrometers and diameters between 0.3 and 5.0 micrometers. The axons contain neurofilaments and mitochondria, although no axolemma or neurotubules are evident. The preparation contains cerebroside and sulfatide, yet is essentially free of myelin.