The perforating branches (PFB's) of the middle cerebral artery (MCA) were studied in 34 unfixed brain hemispheres which were injected with a polyester resin and dissected under the operating microscope. Five hundred and eight vessels were identified and their site of origin, branching pattern, outer diameter (OD), and length recorded. Four hundred and two PFB's (79%) originated from the main trunk of the MCA before its division; the remaining 106 vessels (21%) had their origin from branches of the MCA as follows: superior trunk, 43 vessels (8.5%); inferior trunk, 30 vessels (6%); middle trunk, four vessels (0.8%); early temporal branch, 27 vessels (5.3%); and early frontal branch, two vessels (0.4%). The number of PFB's in each hemisphere varied from five to 29 (mean 14.9 +/- 0.7 vessels). The great majority of PFB's (96%) originated along the proximal 17 mm of the MCA. The PFB's arising in the first 10 mm had a mean OD of 0.35 +/- 0.01 mm and a mean length of 9.25 +/- 0.19 mm, and those arising from the second 10 mm had a mean OD of 0.47 +/- 0.02 mm and a mean length of 16.67 +/- 1.4 mm. A clear distinction between a medial and lateral group of PFB's was present in only 14 hemispheres (41%). In nine hemispheres (26%), perforating vessels from the anterior cerebral artery (A1 segment) and from the recurrent artery of Heubner replaced the medial group of PFB's of the MCA. In one case this group originated in an accessory MCA. In three hemispheres (9%) a small anastomosis (OD 0.2 mm) was seen between a PFB of the recurrent artery of Heubner and one of the MCA. From a total of 508 PFB's, 255 vessels (50%) originated as single vessels, while 253 vessels (50%) originated as branches of common stems. The OD of the single vessels ranged from 0.1 mm to 1.1 mm (mean 0.39 +/- 0.02 mm), and the length from 3 to 20 mm (mean 10.8 +/- 0.2 mm). The common stems ranged in OD from 0.6 to 1.8 mm (mean 0.87 +/- 0.04 mm), and in length from 1 to 15 mm (mean 4.1 +/- 0.4 mm). The clinical application of these anatomical data to the management of aneurysms and arteriovenous malformations of the MCA, and in the field of interventional neuroradiology is described. The most frequent pathological entities involving the perforating vessels are also discussed.