Objective To establish the spatial course of distal tubule and afferent arterioles after macula densa, and to locate and detect the proteins in the adjacent parts by using three-dimensional visualization technology of microstructure.   Methods C57 BL/6J mice were fixed by perfusion and embedded in epon 812. Tissue blocks were cut perpendicular to the longitudinal axis of the kidney. And a total of 720, 2.5 μm-thick consecutive sections were obtained from the renal capsule to the outer stripe of the renal outer medulla. After aligning the digital microscopic images through computer registration procedures, the tubules and vessels were traced by 3D reconstruction program edited by C Language. Selecting the tissue sections of the contact site and applying the improved immunoperoxidase staining method  to detect H+-ATPase and Pendrin.   Results By reconstructing the course of the afferent arterioles, distal tubules and their extended connecting tubules of 90 nephrons in three mice, analyzing the adjacent relationship and measuring the length, it was found that the close contact between tubules and afferent arterioles mostly occurred in superficial and midcortical nephrons, and the cells at the contact expressed H+-ATPase and Pendrin, which was non-A-non-B intercalated cells.   Conclusion The contact between the tubules after macula densa and the afferent arterioles is not accidental, but is occurred at the non-A-non-B intercalated cells in the end of the distal convoluted tubules or (and) the connecting tubules, suggesting that the contact at this site may antagonize the tubuloglomerular feedback at the macula densa, thereby restoring glomerular filtration rate and renal blood flow.