A new approach was developed to minimize inevitable damage to nervous and meningeal tissue due to implantation of a sampling tube allowing multiple withdrawal of cerebrospinal fluid (CSF) from the cisterna magna in adult rats. A tube was secured on the atlanto-occipital membrane. Thereafter, a hole was cut through the membrane, allowing flow of CSF from the cisterna magna to the tube. CSF could be sampled repeatedly for at least 1 week. There was no blood-brain barrier damage. The pressure in the cisterna magna remained normal as did the estimated rate of CSF formation. Very few blood cells contaminated the CSF. There was very little evidence of inflammation. The nervous tissue was undamaged as shown by exclusion of a dye-protein complex. The CSF concentrations of the cytosolic neuronal protein neuron-specific enolase (NSE), and of the astrocyte protein S-100 were very low. The pattern of amino acids remained within normal limits. Scanning electron microscopy revealed that clot and reactive changes were restricted to the vicinity of the connecting hole. We conclude that our approach to positioning a tube on the atlanto-occipital membrane and then connecting it to the cisterna magna reproducibly and reliably enables 'atraumatic' multiple sampling of CSF.