It is accepted that the ionic composition of the medium perfused through a microdialysis probe should match that of the extracellular fluid (ECF) under physiological conditions. In contrast, the possibility that control artificial cerebrospinal fluid may influence the experimental or pathological conditions under study, by buffering changes in the ECF composition, has been neglected. Spreading depression (SD) is a propagating transient suppression of electrical activity due to cellular depolarization which may contribute to neuronal damage in focal ischaemia, and underlie the migraine aura Here we report that microdialysis markedly inhibits SD propagation, by buffering the sudden increase in extracellular K+ associated with this event. This effect is independent of the microdialysis flow rate and does not result from tissue injury following probe implantation. This finding clearly illustrates that microdialysis can influence the pathological conditions under investigation.