A simple method for perfusion fixation of avian liver for electron microscopy

Abstract

A simple method and apparatus are described for perfusion fixation of avian liver for electron microscopy. A constant perfusion pressure is maintained at or below a fixed value with the use of a compressed air cylinder and without the use of automatic devices. A hyperosmotic (580–600 mosm) fixative solution containing 4% glutaraldehyde, 0.0005 M CaCl2, and 0.0005 M MgCl2 in 0.05 M phosphate buffer (pH 7.4, buffer osmolality 122 mosm) produced consistent fixation without swelling or undue shrinkage. The cytoplasmic organelles were well preserved; notably, the mitochondria had electron-dense matrices and well-defined cristae. A pressure of 60 mmHg (1 mmHg = 133.322 Pa) maintained by compressed air permits a minimum to maximum flow rate of 14–19 mL∙min−1∙kg body weight−1, and optimum preservations of the architecture of sinusoids.