A technique for improved fixation of the pigeon central nervous system for electron microscopy

The central nervous system (CNS) of the pigeon has been difficult to fix with consistency, and consequently this problem has impeded ultrastructural studies of various parts of the pigeon brain. Here we describe a method for effective fixation of the pigeon CNS and discuss the three principal problems associated with good fixation of this animal's brain. The animal was deeply anesthetized and the thoracic cavity was opened without collapsing the pectoral girdle upon the brachiocephalic trunks and the common carotids. The perfusion pressure was raised to 140–150 mm Hg to overcome the high resistance of the small diameter, long common carotids. Heparin was added to the wash buffer to retard coagulation of blood in the vascular bed of the brain. The method is not foolproof, but with care excellent fixation can be achieved.