A detailed description is given of the distribution of Timm's staining in the human hippocampal formation. In sections cut transverse to the long axis of the hippocampal formation the same pattern of staining is seen throughout almost all of the structure, i.e., through all the main body except the most caudal region and, following the medial bend made by the long axis, through most of the pes hippocampi. The staining typically filled almost the whole of the hilus of the dentate gyrus and the whole depth of the adjacent stratum pyramidale and stratum lucidum of subfield CA3. Rostrally, staining was traced into the tip of the uncus. Caudally, staining was traced to the inferior surface of the splenium at the apex of the caudal taper of the hippocampus. No staining was found in the gyrus fasciolaris, indusium griseum, or anterior hippocampal rudiment. Except in the medial part of the pes hippocampi, the Timm's staining did not reach the proximal border of the subfield CA1. In some sections of the main body of the hippocampus a narrow, infragranular stain-free zone was observed in the hilus. In the dentate gyrus of three older subjects, but not in those of three younger subjects, supragranular staining was found. It is argued that the Timm's staining described in this study is specific to the hippocampal mossy fibres. The distribution of the staining is discussed in relation to the boundaries of subfields CA3 and CA2 as delineated by other authors.